Antipicornaviral compounds, compositions containing them, and methods for their use

ABSTRACT

A magnetic circuit component having a plurality of claws arranged in a plurality of rows, with the base of each claw connected to a common yoke. A plurality of non-interlaced coils constituting a multi-phase winding are included, with the coils being wound around the bases of corresponding claws, and being distributed uniformly in the direction of motion.

RELATED APPLICATIONS

[0001] This application is a division of U.S. patent application Ser.No. 09/947,381, filed on Sep. 7, 2001, which was a division of U.S.patent application Ser. No. 09/421,560, filed on Oct. 20, 1999, now U.S.Pat. No. 6,331,554, which is a division of U.S. patent application Ser.No. 08/991,282, filed Dec. 16, 1997, now U.S. Pat. No. 6,030,371, whichis a continuation-in-part of U.S. patent application Ser. No.08/825,331, filed Mar. 28, 1997, now abandoned.

[0002] Additionally, this application relates to U.S. patent applicationSer. No. 08/850,398, filed May 2, 1997, now U.S. Pat. No. 5,856,530; toU.S. patent application Ser. No. 08/991,739, filed Dec. 16, 1997, nowU.S. Pat. No. 5,962,487; and to U.S. Provisional Application No.60/046,204, filed May 12, 1997. Each of these applications are entirelyincorporated herein by reference.

BACKGROUND OF THE INVENTION

[0003] The invention pertains to the discovery and use of new compoundsthat inhibit the enzymatic activity of picornaviral 3C proteases,specifically rhinovirus proteases (RVPs), as well as retard viral growthin cell culture.

[0004] The picornaviruses are a family of tiny non-enveloped positivestranded RNA containing viruses that infect humans and other animals.These viruses include the human rhinoviruses, human polioviruses, humancoxsackieviruses, human echoviruses, human and bovine enteroviruses,encephalomyocarditis viruses, menigovirus, foot and mouth viruses,hepatitis A virus and others. The human rhinoviruses are a major causeof the common cold. To date, there are no effective therapies to curethe common cold, only treatments that relieve the symptoms.

[0005] Picornaviral infections may be treated by inhibiting theproteolytic 3C enzymes. These enzymes are required for the naturalmaturation of the picornaviruses. They are responsible for theautocatalytic cleavage of the genomic, large polyprotein into theessential viral proteins. Members of the 3C protease family are cysteineproteases, where the sulfhydryl group most often cleaves theglutamine-glycine amide bond. Inhibition of 3C proteases is believed toblock proteolytic cleavage of the polyprotein, which in turn can retardthe maturation and replication of the viruses by interfering with viralparticle production. Therefore, inhibiting the processing of thiscysteine protease with selective, small molecules that are specificallyrecognized should represent an important and useful approach to treatand cure viral infections of this nature and, in particular, the commoncold.

SUMMARY OF THE INVENTION

[0006] The present invention is directed to compounds that function aspicomaviral 3C protease inhibitors, particularly those that haveantiviral activity. It is further directed to the use of such 3Cprotease inhibitors. The Inventors demonstrate that the compounds of thepresent invention bind to rhinovirus 3C proteases and preferably haveantiviral cell culture activity. The enzymatic inhibition assays usedreveal that these compounds can bind irreversibly, and the cell cultureassays demonstrate that these compounds can possess antiviral activity.

[0007] The present invention is directed to compounds of the formula(I):

[0008] wherein

[0009] M is O or S;

[0010] R₁ is H, F, an alkyl group, OH, SH, or an O-alkyl group;

[0011] R₂ and R₅ are independently selected from H,

[0012] or an alkyl group, wherein the alkyl group is different from

[0013] with the proviso that at least one of R₂ or R₅ must be

[0014] and wherein, when R₂ or R₅ is

[0015] X is ═CH or ═CF and Y₁ is ═CH or ═CF,

[0016] or X and Y₁ together with Q′ form a three-membered ring in whichQ′ is —C(R₁₀)(R₁₁)— or —O—, X is —CH— or —CF—, and Y₁ is —CH—, —CF—, or—C(alkyl)-, where R₁₀ and R₁₁ independently are H, a halogen, or analkyl group, or, together with the carbon atom to which they areattached, form a cycloalkyl group or a heterocycloalkyl group,

[0017] or X is —CH₂—, —CF₂—, —CHF—, or —S—, and Y₁ is —O—, —S—, —NR₂—,—C(R₁₃)(R₁₄)—, —C(O)—, —C(S)—, or —C(CR₁₃R₁₄)—,

[0018] wherein R₁₂ is H or alkyl, and R₁₃ and R₁₄ independently are H,F, or an alkyl group, or, together with the atoms to which they arebonded, form a cycloalkyl group or a heterocycloalkyl group;

[0019] A₁ is C, CH, CF, S, P, Se, N, NR₁₅, S(O), Se(O), P—OR₁₅, orP—NR₁₅R₁₆,

[0020] wherein R₁₅ and R₁₆ independently are an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, or aheteroaryl group, or, together with the atom to which they are bonded,form a heterocycloalkyl group;

[0021] D₁ is a moiety with a lone pair of electrons capable of forming ahydrogen bond; and

[0022] B₁ is H, F, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —OR₁₇, —SR₁₇,—NR₁₇R₁₈, —NR₁₉NR₁₇R₁₈, or —NR₁₇OR₁₈,

[0023] wherein R₁₇, R₁₈, and R₁₉ independently are H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, or an acyl group;

[0024] and with the provisos that when D₁ is the moiety ≡N with a lonepair of electrons capable of forming a hydrogen bond, B₁ does not exist;and when A₁ is an sp³ carbon, B₁ is not —NR₁₇R₁₈ when D₁ is the moiety—NR₂₅R₂₆ with a lone pair of electrons capable of forming a hydrogenbond, wherein R₂₅ and R₂₆ are independently H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, or aheteroaryl group;

[0025] and wherein D₁—A₁—B₁ optionally forms a nitro group where A₁ isN;

[0026] and further wherein, when R₂ or R₅ is

[0027] X is ═CH or ═CF and Y₂ is ═C, ═CH, or ═CF,

[0028] or X and Y₂ together with Q′ form a three-membered ring in whichQ′ is —C(R₁₀)(R₁₁)— or —O—, X is —CH— or —CF—, and Y₂ is —CH—, —CF—, or—C(alkyl)-, where R₁₀ and R₁₁ independently are H, a halogen, or analkyl group, or, together with the carbon atom to which they areattached, form a cycloalkyl group or a heterocycloalkyl group,

[0029] or X is —CH₂—, —CF₂—, —CHF—, or —S—, and Y₂ is —O—, —S—,—N(R′₁₂)—, —C(O)—, —C(R′₁₃)(R′₁₄)—, —C(S)—, or —C(CR′₁₃R′₁₄)—,

[0030] wherein R′₁₂ is H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —OR′₁₃,—NR′₁₃R′₁₄, —C(O)—R′₁₃, —SO₂R′₁₃, or —C(S)R′₁₃, and R′₁₃ and R′₁₄,independently are H, F, or an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, or a heteroaryl group, or,together with the atom to which they are attached, form a cycloalkylgroup or a heterocycloalkyl group;

[0031] A₂ is C, CH, CF, S, P, Se, N, NR₁₅, S(O), Se(O), P—OR₁₅, orP—NR₁₅R₁₆,

[0032] wherein R₁₅ and R₁₆ independently are an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, or aheteroaryl group or, together with the atom to which they are bonded,form a heterocycloalkyl group;

[0033] D₂ is a moiety with a lone pair of electrons capable of forming ahydrogen bond; and

[0034] B₂ is H, F, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —OR₁₇, —SR₁₇,—NR₁₇R₁₈, —NR₁₉NR₁₇R₁₈, or —NR₁₇OR₁₈,

[0035] wherein R₁₇, R₁₈, and R₁₉ independently are H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, or an acyl group;

[0036] and further wherein any combination of Y₂, A₂, B₂, and D₂ forms acycloalkyl group, a heterocycloalkyl group, an aryl group, or aheteroaryl group;

[0037] R₃ and R₆ are independently H, F, an alkyl group, a cycloalkylgroup, a heterocycloalkyl group, an aryl group, a heteroaryl group,—C(O)R₁₇, —OR₁₇, —SR₁₇, —NR₁₇R₁₈, —NR₁₉NR₁₇R₁₈, or —NR₁₇OR₁₈,

[0038] wherein R₁₇, R₁₈, and R₁₉ independently are H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, or an acyl group;

[0039] or, R₃ and R₆, together with the carbon atom to which they areattached, form a cycloalkyl group or a heterocycloalkyl group;

[0040] R₇ is H, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, a heteroaryl group, —OR₁₇, —SR₁₇, —NR₁₇R₁₈,—NR₁₉NR₁₇R₁₈, or —NR₁₇OR₁₈,

[0041] wherein R₁₇, R₁₈, and R₁₉ independently are H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, or an acyl group;

[0042] or R₇, together with R₃ or R₆ and the atoms to which they areattached, form a heterocycloalkyl group;

[0043] R₂₀ is H, OH, or any suitable organic moiety; and

[0044] Z and Z₁ are independently H, F, an alkyl group, a cycloalkylgroup, a heterocycloalkyl group, an aryl group, a heteroaryl group,—C(O)R₂₁, —CO₂R₂₁, —CN, —C(O)NR₂₁R₂₂, —C(O)NR₂₁OR₂₂, —C(S)R₂₁,—C(S)NR₂₁R₂₂, —NO₂, —SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂, —SO(NR₂₁)(OR₂₂),—SONR₂₁, —SO₃R₂₁, —PO(OR₂₁)₂, —PO(R₂₁)(R₂₂), —PO(NR₂₁R₂₂)(OR₂₃),—PO(NR₂₁R₂₂)(NR₂₃R₂₄), —C(O)NR₂₁NR₂₂R₂₃, or —C(S)NR₂₁NR₂₂R₂₃,

[0045] wherein R₂₁, R₂₂, R₂₃, and R₂₄ are independently H, an alkylgroup, a cycloalkyl group, a heterocycloalkyl group, an aryl group, aheteroaryl group, an acyl group, or a thioacyl group, or wherein any twoof R₂₁, R₂₂, R₂₃, and R₂₄, together with the atom(s) to which they arebonded, form a heterocycloalkyl group;

[0046] or Z₁, as defined above, together with R₁, as defined above, andthe atoms to which Z₁ and R₁ are bonded, form a cycloalkyl orheterocycloalkyl group,

[0047] or Z and Z₁, both as defined above, together with the atoms towhich they are bonded, form a cycloalkyl or heterocycloalkyl group;

[0048] and pharmaceutically acceptable prodrugs, salts, activemetabolites, and solvates thereof,

[0049] and wherein these compounds, pharmaceutically acceptableprodrugs, salts, active metabolites, and solvates preferably haveantipicomaviral activity with an EC₅₀ less than or equal to 10 μM in theHI-HeLa cell culture assay, and more preferably antirhinoviral activitywith an EC₅₀ less than or equal to 10 μM in the HI-HeLa cell cultureassay and/or anticoxsachieviral activity with an EC₅₀ less than or equalto 10 μM in the HI-HeLa cell culture assay.

DETAILED DESCRIPTION OF THE INVENTION

[0050] The present invention relates to compounds of the formula I:

[0051] wherein R₁, R₂, R₃, R₅, R₆, R₇, R₂₀, M, Z, and Z₁ are as definedabove, and to the pharmaceutically acceptable prodrugs, salts, activemetabolites, and solvates thereof, where these compounds,pharmaceutically acceptable prodrugs, salts, active metabolites, andsolvates preferably have antipicornaviral activity with an EC₅₀ lessthan or equal to 10 μM in the HI-HeLa cell culture assay, and morepreferably antirhinoviral activity with an EC₅₀ less than or equal to 10μM in the HI-HeLa cell culture assay and/or anticoxsachieviral activitywith an EC₅₀ less than or equal to 10 μM in the HI-HeLa cell cultureassay.

[0052] The present invention preferably relates to compounds of theformula X:

[0053] wherein

[0054] R₆₁ is H, F or an alkyl group;

[0055] R₆₂ is selected from one of the following moieties:

[0056] wherein

[0057] R₃₅ is H, an alkyl group, an aryl group, —OR₃₈, or —NR₃₈R₃₉,

[0058] wherein R₃₈ and R₃₉ independently are H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, or an acyl group; and

[0059] R₃₆ is H or an alkyl group, or R₃₅ and R₃₆, together with thenitrogen atom to which they are attached, form a heterocycloalkyl groupor a heteroaryl group;

[0060] R₃₇ is an alkyl group, an aryl group, or —NR₃₈R₃₉, wherein R₃₈and R₃₉ are as defined above;

[0061] R₅₀ is H, an alkyl group, an aryl group, —OR₃₈, —SR₃₉, —NR₃₈R₃₉,—NR₄₀NR₃₈R₃₉, or —NR₃₈OR₃₉, or R₅₀ and R₃₆, together with the atoms towhich they are attached, form a heterocycloalkyl group;

[0062] wherein R₃₈ and R₃₉ are as defined above and R₄₀ is H, an alkylgroup, a cycloalkyl group, a heterocycloalkyl group, an aryl group, aheteroaryl group, or an acyl group; and

[0063] n is 0, 1, or 2;

[0064] R₆₃ is H or an alkyl group;

[0065] R₆₄ is H, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, or a heteroaryl group;

[0066] R₆₅ is H or an alkyl group;

[0067] R₆₆ is H, an acyl group, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a sulfonyl group, or a heteroarylgroup;

[0068] R₆₇ is H or an alkyl group; and

[0069] Z and Z₁ are independently H, F, an alkyl group, a cycloalkylgroup, a heterocycloalkyl group, an aryl group, a heteroaryl group,—C(O)R₂₁, —CO₂R₂₁, —CN, —C(O)NR₂₁R₂₂, —C(O)NR₂₁OR₂₂, —C(S)R₂₁,—C(S)NR₂₁R₂₂, —NO₂, —SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂, —SO(NR₂₁)(OR₂₂),—SONR₂₁, —SO₃R₂₁, —PO(OR₂₁)₂, —PO(R₂₁)(R₂₂), —PO(NR₂₁R₂₂)(OR₂₃),—PO(NR₂₁R₂₂)(NR₂₃R₂₄), —C(O)NR₂₁NR₂₂R₂₃, or —C(S)NR₂₁NR₂₂R₂₃,

[0070] wherein R₂₁, R₂₂, R₂₃, and R₂₄ are independently H, an alkylgroup, a cycloalkyl group, a heterocycloalkyl group, an aryl group, aheteroaryl group, an acyl group, or a thioacyl group, or wherein any twoof R₂₁, R₂₂, R₂₃, and R₂₄, together with the atom(s) to which they arebonded, form a heterocycloalkyl group,

[0071] or Z and Z₁, both as defined above, together with the atoms towhich they are attached, form a heterocycloalkyl group;

[0072] and pharmaceutically acceptable prodrugs, salts, activemetabolites, and solvates thereof.

[0073] As used in the present application, the following definitionsapply:

[0074] An “alkyl group” is intended to mean a straight or branched chainmonovalent radical of saturated and/or unsaturated carbon atoms andhydrogen atoms, such as methyl, ethyl, propyl, isopropyl, butyl,isobutyl, t-butyl, ethenyl, pentenyl, butenyl, propenyl, ethynyl,butynyl, propynyl, pentynl, hexynyl, and the like, which may beunsubstituted (i.e., containing only carbon and hydrogen) or substitutedby one or more suitable substituents as defined below.

[0075] A “cycloalkyl group” is intended to mean a non-aromatic,monovalent monocyclic, bicyclic, or tricyclic radical containing 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbon ring atoms, each of whichmay be saturated or unsaturated, and which may be unsubstituted orsubstituted by one or more suitable substituents as defined below, andto which may be fused one or more heterocycloalkyl groups, aryl groups,or heteroaryl groups, which themselves may be unsubstituted orsubstituted by one or more suitable substituents.

[0076] Illustrative examples of cycloalkyl groups include, but are notlimited to, the following moieties:

[0077] A “heterocycloalkyl group” is intended to mean a non-aromatic,monovalent monocyclic, bicyclic, or tricyclic radical, which issaturated or unsaturated, containing 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, or 18 ring atoms, and which includes 1, 2, 3, 4, or5 heteroatoms selected from nitrogen, oxygen, and sulfur, wherein theradical is unsubstituted or substituted by one or more suitablesubstituents as defined below, and to which may be fused one or morecycloalkyl groups, aryl groups, or heteroaryl groups, which themselvesmay be unsubstituted or substituted by one or more suitablesubstituents. Illustrative examples of heterocycloalkyl groups include,but are not limited to the following moieties:

[0078] An “aryl group” is intended to mean an aromatic, monovalentmonocyclic, bicyclic, or tricyclic radical containing 6, 10, 14, 18carbon ring atoms, which may be unsubstituted or substituted by one ormore suitable substituents as defined below, and to which may be fusedone or more cycloalkyl groups, heterocycloalkyl groups, or heteroarylgroups, which themselves may be unsubstituted or substituted by one ormore suitable substituents. Illustrative examples of aryl groupsinclude, but are not limited to, the following moieties:

[0079] A “heteroaryl group” is intended to mean an aromatic monovalentmonocyclic, bicyclic, or tricyclic radical containing 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, or 18 ring atoms, including 1, 2, 3, 4,or 5 heteroatoms selected from nitrogen, oxygen, and sulfur, which maybe unsubstituted or substituted by one or more suitable substituents asdefined below, and to which may be fused one or more cycloalkyl groups,heterocycloalkyl groups, or aryl groups, which themselves may beunsubstituted or substituted by one or more suitable substituents.Illustrative examples of heteroaryl groups include, but are not limitedto, the following moieties:

[0080] An “acyl group” is intended to mean a —C(O)—R radical, wherein Ris any suitable substituent as defined below.

[0081] A “thioacyl group” is intended to mean a —C(S)—R radical, whereinR is any suitable substituent as defined below.

[0082] A “sulfonyl group” is intended to mean a —SO₂R radical, wherein Ris any suitable substituent as defined below.

[0083] The term “suitable substituent” is intended to mean any of thesubstituents recognizable, such as by routine testing, to those skilledin the art as not adversely affecting the inhibitory activity of theinventive compounds. Illustrative examples of suitable substituentsinclude, but are not limited to, hydroxy groups, oxo groups, alkylgroups, acyl groups, sulfonyl groups, mercapto groups, alkylthio groups,alkoxy groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups,heteroaryl groups, carboxy groups, amino groups, alkylamino groups,dialkylamino groups, carbamoyl groups, aryloxy groups, heteroarlyoxygroups, arylthio groups, heteroarylthio groups, and the like.

[0084] The term “suitable organic moiety” is intended to mean anyorganic moiety recognizable, such as by routine testing, to thoseskilled in the art as not adversely affecting the inhibitory activity ofthe inventive compounds. Illustrative examples of suitable organicmoieties include, but are not limited to, hydroxy groups, alkyl groups,oxo groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups,heteroaryl groups, acyl groups, sulfonyl groups, mercapto groups,alkylthio groups, alkoxy groups, carboxy groups, amino groups,alkylamino groups, dialkylamino groups, carbamoyl groups, arylthiogroups, heteroarylthio groups, and the like.

[0085] A “hydroxy group” is intended to mean the radical —OH.

[0086] An “amino group” is intended to mean the radical —NH₂.

[0087] An “alkylamino group” is intended to mean the radical —NHR whereR is an alkyl group as defined above.

[0088] A “dialkylamino group” is intended to mean the radical—NR_(a)R_(b) where R_(a) and R_(b) are each independently an alkyl groupas defined above.

[0089] An “alkoxy group” is intended to mean the radical —OR where R isan alkyl group as defined above, for example, methoxy, ethoxy, propoxy,and the like.

[0090] An “alkoxycarbonyl group” is intended to mean the radical —C(O)ORwhere R is an alkyl group as defined above.

[0091] An “alkylsulfonyl group” is intended to mean the radical —SO₂Rwhere R is an alkyl group as defined above.

[0092] An “alkylaminocarbonyl group” is intended to mean the radical—C(O)NHR where R is an alkyl group as defined above.

[0093] A “dialkylaminocarbonyl group” is intended to mean the radical—C(O)NR_(a)R_(b) where R_(a) and R_(b) are each independently an alkylgroup as defined above.

[0094] A “mercapto group” is intended to mean the radical —SH.

[0095] An “alkylthio group” is intended to mean the radical —SR where Ris an alkyl group as defined above.

[0096] A “carboxy group” is intended to mean the radical —C(O)OH.

[0097] A “carbamoyl group” is intended to mean the radical —C(O)NH₂.

[0098] An “aryloxy group” is intended to mean the radical —OR_(c) whereR_(c) is an aryl group as defined above.

[0099] A “heteroaryloxy group” is intended to mean the radical —OR_(d)where R_(d) is a heteroaryl group as defined above.

[0100] An “arylthio group” is intended to mean the radical —SR_(c) whereR_(c) is an aryl group as defined above.

[0101] A “heteroarylthio group” is intended to mean the radical —SR_(d)where R_(d) is a heteroaryl group as defined above.

[0102] A “pharmaceutically acceptable prodrug” is intended to mean acompound that may be converted under physiological conditions or bysolvolysis to a compound of formula I or formula X.

[0103] A “pharmaceutically acceptable active metabolite” is intended tomean a pharmacologically active product produced through metabolism inthe body of a compound of formula I or formula X.

[0104] A “pharmaceutically acceptable solvate” is intended to mean asolvate that retains the biological effectiveness and properties of thebiologically active components of compounds of formulas I and X.Examples of pharmaceutically acceptable solvates include, but are notlimited to, compounds of formula I or X in combination with water,isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, orethanolamine.

[0105] A “pharmaceutically acceptable salt” is intended to mean a saltthat retains the biological effectiveness and properties of the freeacids and bases of compounds of formulas I and X and that is notbiologically or otherwise undesirable. Examples of pharmaceuticallyacceptable salts include, but are not limited to, sulfates,pyrosulfates, bisulfates, sulfites, bisulfites, phosphates,monohydrogenphosphates, dihydrogenphosphates, metaphosphates,pyrophosphates, chlorides, bromides, iodides, acetates, propionates,decanoates, caprylates, acrylates, formates, isobutyrates, caproates,heptanoates, propiolates, oxalates, malonates, succinates, suberates,sebacates, fumarates, maleates, butyne-1,4-dioates, hexyne-1,6-dioates,benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates,hydroxybenzoates, methoxyenzoates, phthalates, sulfonates,xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates,citrates, lactates, γ-hydroxybutyrates, glycollates, tartrates,methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates,naphthalene-2-sulfonates, and mandelates.

[0106] If the inventive compound is a base, the desired salt may beprepared by any suitable method known to the art, including treatment ofthe free base with an inorganic acid, such as hydrochloric acid;hydrobromic acid; sulfiric acid; nitric acid; phosphoric acid; and thelike, or with an organic acid, such as acetic acid; maleic acid;succinic acid; mandelic acid; fumaric acid; malonic acid; pyruvic acid;oxalic acid; glycolic acid; salicylic acid; pyranosidyl acids such asglucuronic acid and galacturonic acid; alpha-hydroxy acids such ascitric acid and tartaric acid; amino acids such as aspartic acid andglutamic acid; aromatic acids such as benzoic acid and cinnamic acid;sulfonic acids such as p-toluenesulfonic acid or ethanesulfonic acid; orthe like.

[0107] As generally understood by those skilled in the art, an opticallypure compound is one that is enantiomerically pure. As used herein, theterm “optically pure” is intended to mean a compound which comprises atleast a sufficient amount of a single enantiomer to yield a compoundhaving the desired pharmacological activity. Preferably, “opticallypure” is intended to mean a compound that comprises at least 90% of asingle isomer (80% enantiomeric excess), preferably at least 95% (90%e.e.), more preferably at least 97.5% (95% e.e.), and most preferably atleast 99% (98% e.e.).

[0108] Preferably in the above formulas I and X, R₁ and R₆₁, are H or F.In the compounds of formula I, preferably M is O.

[0109] Preferably R₂₀ in formula I is H, an alkyl group, a cycloalkylgroup, a heterocycloalkyl group, an aryl group, a heteroaryl group,—OR₁₇, —SR₁₇, —NR₁₇R₁₈, —NR₁₉NR₁₇R₁₈, or —NR₁₇OR₁₈, wherein R₁₇, R₁₈,and R₁₉ independently are H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, or an acylgroup. More preferably R₂₀ is the alkyl group —C(R₄₁)(R₄₂)NR₄₃R₄₄,wherein R₄₁, and R₄₂ independently are H, an alkyl group, a cycloalkylgroup, a heterocycloalkyl group, an aryl group, or a heteroaryl group;and R₄₃ and R₄₄ independently are H, an alkyl group, a cycloalkyl group,a heterocycloalkyl group, an aryl group, a heteroaryl group, —NR₄₅R₄₆,—C(O)R₄₅, —C(S)R₄₅, —C(O)NR₄₅R₄₆, —C(S)NR₄₅R₄₆, —C(O)NR₄₅OR₄₆,—C(S)NR₄₅OR₄₆, —C(O)SR₄₅, —C(O)OR₄₅, —C(S)OR₄₅, —C(S)SR₄₅, —OR₄₅, —SR₄₅,—C(O)NR₄₅NR₄₆R₄₇, —C(S)NR₄₅NR₄₆R₄₇, —SOR₄₅, —SO₂R₄₅, —S(O)NR₄₅R₄₆,—S(O)NR₄₅(OR₄₆), —SO₂NR₄₅R₄₆, —SO₂N₄₅(OR₄₆), or —SO₃R₄₅, wherein R₄₅,R₄₆, and R₄₇ independently are H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, or whereinany combination of R₄₁, R₄₂, R₄₃, and R₄₄, together with the atoms towhich they are attached, form a cycloalkyl group or a heterocycloalkylgroup. Preferably at least one of R₄₃ and R₄₄ is —C(O)SR₄₅ or —C(O)OR₄₅.Preferably R₄₅ is an alkyl group, a cycloalkyl group, an aryl group, aheterocycloalkyl group, or a heteroaryl group, and more preferably aC₁-C₁₀ alkyl group.

[0110] In the compounds of formula I, preferably D₁ and D₂ are —OR₂₅,═O, ═S, ≡N, ═NR₂₅, or —NR₂₅R₂₆, wherein R₂₅ and R₂₆ are independently H,an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, or a heteroaryl group, or, together with the nitrogen atom towhich they are bonded, form a heterocycloalkyl group, and morepreferably D₁ and D₂ are ═O. Preferably, in the compounds of formula I,A₁ and A₂ are C, CH, S, or S(O), and more preferably A₁ and A₂ are C.

[0111] Preferably, in the compounds of formula I, B₁ and B₂ are NR₁₇R₁₈,wherein R₁₇ and R₁₈ are independently H, an alkyl group, a cycloalkylgroup, a heterocycloalkyl group, an aryl group, a heteroaryl group, oran acyl group.

[0112] In the compounds of formula I and X, preferably Z and Z₁ areindependently H, an aryl group, or a heteroaryl group, —C(O)R₂₁,—CO₂R₂₁, —CN, —C(O)NR₂₁,R₂₂, —C(O)NR₂₁OR₂₂, —C(S)R₂₁, —C(S)NR₂₁R₂₂,—NO₂, —SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂, —SO(NR₂₁)(OR₂₂), —SONR₂₁, —SO₃R₂₁,—C(O)NR₂₁NR₂₂R₂₃, or —C(S)NR₂₁NR₂₂R₂₃, wherein R₂₁, R₂₂, and R₂₃ areindependently H, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, a heteroaryl group, an acyl group, or a thioacylgroup, or wherein any two of R₂₁, R₂₂, and R₂₃, together with theatom(s) to which they are bonded, form a heterocycloalkyl group, or Zand Z₁, both as defined above, together with the atoms to which they areattached, form a heterocycloalkyl group.

[0113] In the compounds of formula X, preferably R₆₆ is —C(O)OR₆₈ or—C(O)SR₆₈, wherein R₆₈ is an alkyl group, a cycloalkyl group, an arylgroup, a heterocycloalkyl group, or a heteroaryl group

[0114] Particularly preferred embodiments of the present inventioninclude the following compounds (1-8, 10, and 11) of formula II:

[0115] wherein R₁, R₅, R₆, R₇, R₄₂, R₄₃, and Z are H, R₂ isCH₂CH₂C(O)NH₂, and

[0116] (1) R₃ is CH₂Ph, R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

[0117] (2) R₃ is CH₂Ph, R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

[0118] (3) R₃ is CH₂Ph, R₄₁ is CH₂CH(CH₃)₂, Z₁ is

[0119]  and R₄₄ is

[0120] (4) R₃ is CH₂Ph, R₄₁ is CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

[0121] (5) R₃ is CH₂Ph, R₄₁ is CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

[0122] (6) R₃ is CH₂Ph, R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

[0123] (7) R₃ is CH₂Ph, R₄₁ is CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

[0124] (8) R₃ is CH₂Ph, R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

[0125] (10) R₃ is CH₂(p-CH₃)Ph, R₄₁ is CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, andR₄₄ is

[0126] (11) R₃ is CH₂(p-CH₃)Ph, R₄₁ is CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, andR₄₄ is

[0127] and a compound (9) of formula III:

[0128] wherein R₁, R₅, R₆, R₇, R₄₂, R₄₃, and Z are H, R₃ is CH₂Ph, R₂ isCH₂CH₂C(O)NH₂,

[0129] R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

[0130]  and pharmaceutically acceptable prodrugs, salts, activemetabolites, and solvates thereof.

[0131] The present invention is still further directed to compositionscomprising at least one compound of formula II:

[0132] wherein R₁, R₅, R₆, R₇, R₄₂, R₄₃, and Z are H, R₃ is CH₂Ph, R₂ isCH₂CH₂C(O)NH₂,

[0133] R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

[0134]  or a pharmaceutically acceptable prodrug, salt, activemetabolite, or solvate thereof,

[0135] and at least one compound of formula III:

[0136] wherein R₁, R₅, R₆, R₇, R₄₂, R₄₃, and Z are H, R₃ is CH₂Ph, R₂ isCH₂CH₂C(O)NH₂,

[0137] R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

[0138]  or a pharmaceutically acceptable prodrug, salt, activemetabolite, or solvate thereof.

[0139] Additional preferred compounds according to the present inventioninclude the following compounds (12 through 25) of formula IV:

[0140] wherein R₁, R₅, R₆, R₇, and R₄₂ are H, R₂ is CH₂CH₂C(O)NH₂, and

[0141] (12) R₃ is CH₂(p-CH₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH₂Ph,and R₄₄ is

[0142] (13) R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH(CH₃)₂,and R₄₄ is

[0143] (14) R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH(CH₃)₂,and R₄₄ is

[0144] (15) R₃ is CH₂(p-CF₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ isCH(CH₃)₂, and R₄₄ is

[0145] (16) R₃ is CH₂(p-CF₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ isCH(CH₃)₂, and R₄₄ is

[0146] (17) R₃ is CH₂(p-CH₃)Ph, Z and Z₁ together form

[0147]  (where * indicates the point of attachment and the carbonylgroup is cis to the R₁ group), R₄₁ is CH(CH₃)₂, and R₄₄ is

[0148] (18) R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH₂Ph, andR₄₄ is

[0149] (19) R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ isCH₂CH(CH₃)₂, and R₄₄ is

[0150] (20) R₃ is CH₂(p-CH₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ isCH(CH₃)₂, and R₄₄ is

[0151] (21) R₃ is CH₂(p-CH₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ isCH₂CH(CH₃)₂, and R₄₄ is

[0152] (22) R₃ is CH2Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is C(CH₃)₃, andR₄₄ is

[0153] (23) R₃ is CH₂(p-CH₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ isCH(CH₃)₂, and R₄₄ is

[0154] (24) R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ iscyclohexyl, and R₄₄ is

[0155] (25) R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH(CH₃)₂,and R₄₄ is

[0156] and pharmaceutically acceptable prodrugs, salts, activemetabolites, or solvates thereof.

[0157] The present invention is even further directed to methods ofinhibiting picornaviral 3C protease activity, comprising contacting theprotease with an effective amount of a compound of formula I or X or apharmaceutically acceptable prodrug, salt, active metabolite, or solvatethereof. For example, one can inhibit picornaviral 3C protease activityin mammalian tissue by administering a compound of formula I or X or apharmaceutically acceptable prodrug, salt, active metabolite, or solvatethereof. More particularly, the present invention is directed to methodsof inhibiting rhinoviral protease activity.

[0158] The activity of the inventive compounds as inhibitors ofpicornaviral 3C protease activity may be measured by any of the methodsavailable to those skilled in the art, including in vivo and in vitroassays. An example of a suitable assay for activity measurements is theAntiviral HI-HeLa Cell Culture Assay, described herein.

[0159] Administration of the compounds of the formula I or X, or theirpharmaceutically acceptable prodrugs, salts, active metabolites, andsolvates, may be performed according to any of the accepted modes ofadministration available to those skilled in the art. Illustrativeexamples of suitable modes of administration include, but are notlimited to, oral, nasal, parenteral, topical, transdermal, and rectal.

[0160] The inventive compounds of formulas I and X, and theirpharmaceutically acceptable prodrugs, salts, active metabolites, andsolvates, may be administered as a pharmaceutical composition in anysuitable pharmaceutical form recognizable to the skilled artisan.Suitable pharmaceutical forms include, but are not limited to, solid,semisolid, liquid, or lyopholized formulations, such as tablets,powders, capsules, suppositories, suspensions, and aerosols. Thepharmaceutical composition may also include suitable excipients,diluents, vehicles, and carriers, as well as other pharmaceuticallyactive agents, depending upon the intended use.

[0161] Acceptable methods of preparing suitable pharmaceutical forms ofthe pharmaceutical compositions are known to those skilled in the art.For example, pharmaceutical preparations may be prepared followingconventional techniques of the pharmaceutical chemist involving stepssuch as mixing, granulating, and compressing when necessary for tabletforms, or mixing, filling, and dissolving the ingredients asappropriate, to give the desired products for oral, parenteral, topical,intravaginal, intranasal, intrabronchial, intraocular, intraaural,and/or rectal administration.

[0162] Solid or liquid pharmaceutically acceptable carriers, diluents,vehicles, or excipients may be employed in the pharmaceuticalcompositions. Illustrative solid carriers include starch, lactose,calcium sulphate dihydrate, terra alba, sucrose, talc, gelatin, pectin,acacia, magnesium stearate, and stearic acid. Illustrative liquidcarriers may include syrup, peanut oil, olive oil, saline solution, andwater. The carrier or diluent may include a suitable prolonged-releasematerial, such as glyceryl monostearate or glyceryl distearate, alone orwith a wax. When a liquid carrier is used, the preparation may be in theform of a syrup, elixir, emulsion, soft gelatin capsule, sterileinjectable liquid (e.g. solution), or a nonaqueous or aqueous liquidsuspension.

[0163] A dose of the pharmaceutical composition contains at least atherapeutically effective amount of the active compound (i.e., acompound of formula I or X or a pharmaceutically acceptable prodrug,salt, active metabolite, or solvate thereof) and preferably is made upof one or more pharmaceutical dosage units. The selected dose may beadministered to a mammal, for example, a human patient, in need oftreatment mediated by inhibition of 3C protease activity, by any knownmethod of administering the dose including topical, for example, as anointment or cream; orally; rectally, for example, as a suppository;parenterally by injection; or continuously by intravaginal, intranasal,intrabronchial, intraaural, or intraocular infusion.

[0164] A “therapeutically effective amount” is intended to mean thatamount of a compound of formula I or X that, when administered to amammal in need thereof, is sufficient to effect treatment for diseaseconditions alleviated by the inhibition of the activity of one or morepicomaviral 3C proteases, such as human rhinoviruses, human poliovirus,human coxsackieviruses, encephalomyocarditis viruses, menigovirus, andhepatitis A virus. The amount of a given compound of formula I or X thatwill correspond to a “therapeutically effective amount” will varydepending upon factors such as the particular compound, the diseasecondition and the severity thereof, the identity of the mammal in needthereof, but it can nevertheless be readily determined by one of skillin the art.

[0165] “Treating” or “treatment” is intended to mean at least themitigation of a disease condition in a mammal, such as a human, that isalleviated by the inhibition of the activity of one or more picomaviral3C proteases, such as human rhinoviruses, human poliovirus, humancoxsackieviruses, encephalomyocarditis viruses, menigovirus, andhepatitis A virus, and includes:

[0166] (a) prophylactic treatment in a mammal, particularly when themammal is found to be predisposed to having the disease condition butnot yet diagnosed as having it;

[0167] (b) inhibiting the disease condition; and/or

[0168] (c) alleviating, in whole or in part, the disease condition.

[0169] The inventive compounds, and their salts, solvates, activemetabolites, and prodrugs, may be prepared by employing the techniquesavailable in the art using starting materials that are readilyavailable. Certain novel and exemplary methods of preparing theinventive compounds are described below.

[0170] Preferably, the inventive compounds of formula I are prepared bythe methods of the present invention, including the general methodsshown below. In each of these general methods, R1, R2, R3, R5, R6, R7,R20, R41, R42, Z, and Z1 are as defined above (for formulae I, II, III,IV, and X).

[0171] General Method I

[0172] In General Method I, amino acid A, where P1 is an appropriateprotecting group for nitrogen, is converted to carbonyl derivative B,where “Lv” is a leaving group. Compound B is subjected to a reactionwhere “Lv” is replaced by R1 to give derivative C. Derivative C is thentransformed into unsaturated product D. Unsaturated compound D isdeprotected to give free amine (or salt thereof) E, or modified one ormore times at R2, R5, Z, and/or Z1 to give one or more modified Dcompounds. Modified D is then deprotected to give amine (or saltthereof) E.

[0173] Amine E is subsequently subjected to an amide-forming reactionwith carboxylic acid F (for which the preparation of representativeexamples is described below) to give final product G. If protectinggroups were used on any R groups (R₁, R₂, R₃, R₅, R₆, R₇, and/or R₂₀),on Z and/or on Z₁, product G is deprotected and/or further modified toyield “deprotected or modified G.”

[0174] In General Method II, amine E, which can be prepared as describedin General Method I, is subjected to an amide-forming reaction withcarboxylic acid H, where P₂ is an appropriate protecting group fornitrogen, and where at least one of R₃ and R₆ is H, to give finalproduct I. Carboxylic acid H can be prepared as a mixture ofdiastereomers as described in Harbeson, S. L., Rich, D. H., J. Med.Chem. 1989, 32, 1378, the disclosure of which is incorporated herein byreference. The P₂ protecting group, along with any additional protectinggroups that were used on any R groups (R₁, R₂, R₃, R₅, R₆, R₇, and/orR₄₁), on Z and/or on Z₁, is subsequently deprotected and/or furthermodified to yield “deprotected or modified I.”

[0175] In General Method III, optically active lactone J, where P₂ is anappropriate protecting group for nitrogen, is transformed by a two-stepprocedure (basic hydrolysis and subsequent oxidation) into carboxylicacid K. Lactone J can be prepared by the method described in GeneralMethod IV below and by many literature methods including, but notlimited to, those described in the following: (a) Herold, P.; Duthaler,R.; Rihs, G.; Angst, C., J. Org. Chem. 1989, 54, 1178; (b) Bradbury, R.H.; Revill, J. M.; Rivett, J. E.; Waterson, D., Tetrahedron Lett. 1989,30, 3845; (c) Bradbury, R. H.; Major, J. S.; Oldham, A. A.; Rivett, J.E.; Roberts, D. A.; Slater, A. M.; Timms, D.; Waterson, D., J. Med.Chem. 1990, 33, 2335; (d) Wuts, P. G.; Ritter, A. R.; Pruitt, L. E., J.Org. Chem. 1992, 57, 6696; (e) Jones, D. M.; Nilsson, B.; Szelke, M., J.Org. Chem. 1993, 58, 2286; (f) Pégorier, L.; Larchevéque, M.,Tetrahedron Lett. 1995, 36, 2753; (g) Dondoni, A.; Perrone, D.; Semola,M. T., J. Org. Chem. 1995, 60, 7927, all of which are incorporatedherein by reference. Carboxylic acid K is not isolated in pure form, butis subjected to an amide-forming reaction with amine E, which can beprepared as described in General Method I, to provide final product L.The P₂ protecting group, along with any additional protecting groupsthat were used on any R groups (R₁, R₂, R₃, R₅, R₇, and/or R₄₁), on Zand/or on Z₁, is subsequently deprotected and/or further modified toyield “deprotected or modified L.”

[0176] Lactone J may be prepared in optically active form by GeneralMethod IV (see: (a) Herold, P.; Duthaler, R.; Rihs, G.; Angst, C., J.Org. Chem. 1989, 54, 1178; (b) Bradbury, R. H.; Revill, J. M.; Rivett,J. E.; Waterson, D., Tetrahedron Lett. 1989, 30, 3845; (c) Bradbury, R.H.; Major, J. S.; Oldham, A. A.; Rivett, J. E.; Roberts, D. A.; Slater,A. M.; Timms, D.; Waterson, D., J. Med. Chem. 1990, 33, 2335). Aγ,δ-unsaturated carboxylic acid M, which incorporates R₄₁, istransformed into the corresponding acid chloride (not shown). This acidchloride is subjected to an amide-forming reaction with a chiral amineor a chiral oxazolidone to provide derivative N (in which X₁ is a chiralamine or a chiral oxazolidone). Compound N is subsequently deprotonated,and the resulting enolate is diastereoselectively alkylated with anelectrophile corresponding to R₃ to provide compound O. This material isthen subjected to a halolactonization reaction to provide halo-lactoneP, in which H₁ is Br or I. Halo-lactone P is subsequently transformedinto azide Q, and this material is then converted into lactone J, whereP₂ is an appropriate protecting group for nitrogen.

[0177] γ,δ-Unsaturated carboxylic acid M may be prepared by GeneralMethod V (see: Herold, P.; Duthaler, R.; Rihs, G.; Angst, C., J. Org.Chem. 1989, 54, 1178). An aldehyde R, which incorporates R₄₁, is coupledwith vinylmagnesium bromide to give alcohol S. Alcohol S is thentransformed into γ,δ-unsaturated carboxylic acid M by a three stepprocedure as follows: (I) treatment with diethyl malonate and catalyticTi(OEt)₄ at 160° C. for 1 hour, (ii) heating at 190° C. for 4 hours, and(iii) hydrolysis with ethanolic KOH at reflux.

[0178] Carboxylic acid K also may be prepared by General Method VI (see:Hoffman, R. V., Tao, J., Tetrahedron, 1997, 53, 7119, which document isentirely incorporated herein by reference). An amino acid T, whichincorporates R₄₁ and where P₂ is an appropriate protecting group fornitrogen, is transformed into β-ketoester U. Compound U is deprotonated,and the resulting anion is condensed with triflate V, which incorporatesR₃. The coupling product thus obtained is treated with trifluoroaceticacid to provide ketoester W, and this material is subsequentlyhydrolyzed to afford carboxylic acid K. Triflate V, in turn, may beprepared from the corresponding alcohol by treatment withtrifluoromethanesulfonic anhydride and 2,6-lutidine.

[0179] Lactone J also may be prepared by General Method VII (see: (a)Askin, D., Wallace, M. A., Vacca, J. P., Reamer, R. A., Volante, R. P.,Shinkai, I. J. Org. Chem. 1992, 57, 2771 (b) McWilliams, J. C.,Armstrong, J. D., Zheng, N., Bhupathy, M., Volante, R. P., Reider, P.J., J. Am. Chem. Soc. 1996, 118, 11970; each of these documents isentirely incorporated herein by reference). An amino acid T, whichincorporates R₄₁ and where P₂ is an appropriate protecting group fornitrogen, is transformed into epoxide X (single diastereomer) by themethod described in: Luly, J. R., Dellaria, J. F., Plattner, J. J.,Soderquist, J. L., Yi, N., J. Org. Chem. 1987, 52, 1487, the disclosureof which is entirely incorporated herein by reference. Alternatively, Xmay be prepared from T as a mixture of diastereomers as described in the“Examples” section of this document. Epoxide X is condensed with theanion derived from compound Y, which incorporates R₃ and in which X₂ isa chiral auxiliary [including (1S,2R)-1-aminoindan-2-ol acetonide] toafford coupling product Z. If X was utilized as a mixture ofdiastereomers, the diastereomer of Z depicted below is purified fromother Z isomers (if any) produced in the coupling reaction. Thismaterial is subsequently cyclized under acidic conditions to providelactone J. Compound Y may be prepared from the corresponding carboxylicacid (not shown) by the method outlined in: Askin, D., Wallace, M. A.,Vacca, J. P., Reamer, R. A., Volante, R. P., Shinkai, I., J. Org. Chem.1992, 57, 2771.

[0180] Suitable protecting groups for nitrogen are recognizable to thoseskilled in the art and include, but are not limited tobenzyloxycarbonyl, t-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,p-methoxybenxyloxycarbonyl, trifluoroacetamide, and p-toluenesulfonyl.Suitable protecting groups for oxygen are recognizable to those skilledin the art and include, but are not limited to —CH₃, —CH₂CH₃, tBu,—CH₂Ph, —CH₂CH═CH₂, —CH₂OCH₂CH₂Si(CH₃)₃, and —CH₂CCl₃. Other examples ofsuitable protecting groups for nitrogen or oxygen can be found in T.Green & P. Wuts, Protective Groups in Organic Synthesis (2nd ed. 1991),the disclosure of which is incorporated herein by reference.

[0181] Suitable leaving groups also are recognizable to those skilled inthe art and include, but are not limited to, Cl, Br, I, sulfonates,O-alkyl groups,

[0182] Other examples of suitable leaving groups are described in J.March, Advanced Organic Chemistry, Reactions, Mechanisms, and Structure(4th ed. 1992) at pages 205, 351-56, 642-43, 647, 652-53, 666, 501,520-21, 569, 579-80, 992-94, 999-1000, 1005, and 1008, the disclosure ofwhich is incorporated herein by reference.

EXAMPLES

[0183] Examples of the processes used to make several of the compoundsof formula I are set forth below. These Examples are intended toillustrate the present invention without limiting it. The structures ofthe compounds of the following examples were confirmed by one or more ofthe following: proton magnetic resonance spectroscopy, infraredspectroscopy, elemental microanalysis, mass spectrometry, thin layerchromatography, melting point, and boiling point.

[0184] Proton magnetic resonance (1H NGR) spectra were determined usinga Varian UNITYplus 300 spectrometer operating at a field strength of 300megahertz (MHZ). Chemical shifts are reported in parts per million (ppm,□) downfield from an internal tetramethylsilane standard. Alternatively,1H NGR spectra were referenced to residual protic solvent signals asfollows: CHCl3=7.26 ppm; DMSO=2.49 ppm, C6HD5=7.15 ppm. Peakmultiplicities are designated as follows: s, singlet; d, doublet; dd,doublet of doublets; t, triplet; q, quartet; br, broad resonance; m,multiplet. Coupling constants (J) are given in hertz (Hz). Infraredabsorption (IR) spectra were obtained using a Perkin-Elmer 1600 seriesFTIR spectrometer. Elemental microanalyses were performed by AtlanticMicrolab Inc., Norcross, Ga. and gave results for the elements statedwithin ±0.4% of the theoretical values.

[0185] Flash column chromatography was performed using Silica gel 60(Merck Art 9385). Analytical thin layer chromatography (TLC) wasperformed using precoated sheets of Silica 60 F254 (Merck Art 5719).Melting points were determined on a Mel-Temp apparatus and areuncorrected. All reactions were performed in septum-sealed flasks undera slight positive pressure of argon unless otherwise noted. Allcommercial reagents were used as received from their respectivesuppliers with the following exceptions. Tetrahydrofuran (THF) wasdistilled from sodium-benzophenone ketyl prior to use. Dichloromethane(CH2Cl2) was distilled from calcium hydride prior to use. Anhydrouslithium chloride was prepared by heating at 110 □C under vacuumovernight. Et2O refers to diethyl ether. DMF refers toN,N-dimethylformamide. DMSO refers to dimethylsulfoxide. Otherabbreviations include: CH3OH (methanol), EtOH (ethanol), EtOAc (ethylacetate), DME (ethylene glycol dimethyl ether), Ac (acetyl), Me(methyl), Et (ethyl), Ph (phenyl), Bn (benzyl), CyPentyl (cyclopentyl),Tr (triphenylmethyl), CBZ (benzyloxycarbonyl), BOC(tert-butoxycarbonyl), Gln (glutamine), Leu (leucine), Phe(phenylalanine), Val (valine). Additionally, “L” represents naturallyoccurring amino acids, “D” represents unnatural amino acids, and “DL”represents a racemic mixture of the two.

[0186] A simplified naming system employing amino acid abbreviations isused to identify some intermediates and final products. When namingcompounds, italicized amino acid abbreviations represent modificationsat the C-terminus of that residue where the following apply:

[0187] 1. acrylic acid esters are reported as “E” (trans) propenoates;

[0188] 2. acrylamides are reported as “E” (trans) propenamides; and

[0189] 3. N-acetyl-pyrrolidin-2-ones are reported as “E” (trans)1-acetyl-3-methylene-pyrrolidin-2-ones.

[0190] In addition, the terminology “AA₁Ψ[COCH₂]-AA₂” indicates that,for any peptide sequence, two amino acids (AA₁ and AA₂) linked by anamide bond are replaced by a ketomethlyene dipeptide isostere moiety.

Example 1

[0191] Preparation of a Mixture of Compound 1 and Compound 9 (˜1:1):Ethyl-3-(CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-Gln)-E-Propenoate.

[0192] Preparation of Intermediate [BOC-L-(Tr-Gln)]-N(OMe)Me.

[0193] Isobutyl chloroformate (4.77 mL, 36.8 mmol, 1.0 equiv) was addedto a solution of [BOC-L-(Tr-Gln)]-OH (18.7 g, 36.7 mmol, 1 equiv) and4-methylmorpholine (8.08 mL, 73.5 mmol, 2.0 equiv) in CH₂Cl₂ (250 mL) at0° C. The reaction mixture was stirred at 0° C. for 20 minutes, thenN,O-dimethylhydroxylamine hydrochloride (3.60 g, 36.7 mmol, 1.0 equiv)was added. The resulting solution was stirred at 0° C. for 20 minutesand at 23° C. for 2 hours, and then was partitioned between water (150mL) and CH₂Cl₂ (2×150 mL). The combined organic layers were dried overNa₂SO₄, and were concentrated. Purification of the residue by flashcolumn chromatography (gradient elution, 40% →20% hexanes in EtOAc)provided [BOC-L-(Tr-Gln)]-N(OMe)Me (16.1 g, 82%) as a white foam:R_(f)=0.22 (50% EtOAc in hexanes); IR (cm⁻¹) 3411, 3329, 3062, 1701,1659; ¹H NGR (CDCl₃) δ 1.42 (s, 9H), 1.63-1.77 (m, 1H), 2.06-2.17 (m,1H), 2.29-2.43 (m, 2H), 3.17 (s, 3H), 3.64 (s, 3H), 4.73 (s, br, 1H),5.38-5.41 (m, 1H), 7.20-7.31 (m, 15H); Anal. (C₃₁H₃₇N₃O₅) C, H, N.

[0194] Preparation of Intermediate [BOC-L-(Tr-Gln)]-H.

[0195] Diisobutylaluminum hydride (50.5 mL of a 1.5 M solution intoluene, 75.8 mmol, 2.5 equiv) was added to a solution of[BOC-L-(Tr-Gln)]-N(OMe)Me (16.1 g, 30.3 mmol, 1 equiv) in THF at −78°C., and the reaction mixture was stirred at −78° C. for 4 hours.Methanol (4 mL) and 1.0 M HCl (10 mL) were added sequentially, and themixture was warmed to 23° C. The resulting suspension was diluted withEt₂O (150 mL) and was washed with 1.0 M HCl (3×100 mL), half-saturatedNaHCO₃ (100 mL), and water (100 mL). The organic layer was dried overMgSO₄, filtered, and concentrated to give crude [BOC-L-(Tr-Gln)]-H (13.8g, 97%) as a white solid: mp=114-116° C.; R_(f)=0.42 (50% EtOAc inhexanes); IR (cm⁻¹) 3313, 1697, 1494; ¹H NGR (CDCl₃) δ 1.44 (s, 9H),1.65-1.75 (m, 1H), 2.17-2.23 (m, 1H), 2.31-2.54 (m, 2H), 4.11 (s, br,1H), 5.38-5.40 (m, 1H), 7.11 (s, 1H), 7.16-7.36 (m, 15H), 9.45 (s, 1H).

[0196] Preparation of IntermediateEthyl-3-[BOC-L-(Tr-Gln)]-E-Propenoate.

[0197] Sodium bis(trimethylsilyl)amide (22.9 mL of a 1.0 M solution inTHF, 22.9 mmol, 1.0 equiv) was added to a solution of triethylphosphonoacetate (5.59 g, 22.9 mmol, 1.0 equiv) in THF (200 mL) at −78°C., and the resulting solution was stirred for 20 minutes at thattemperature. Crude [BOC-L-(Tr-Gln)]-H (10.8 g, 22.9 mmol, 1 equiv) inTHF (50 mL) was added via cannula, and the reaction mixture was stirredfor 2 hours at −78° C., warmed to 0° C. for 10 minutes, and partitionedbetween 0.5 M HCl (150 mL) and a 1:1 mixture of EtOAc and hexanes (2×150mL). The combined organic layers were dried over Na₂SO₄ andconcentrated. Purification of the residue by flash column chromatography(40% EtOAc in hexanes) provided ethyl-3-[BOC-L-(Tr-Gln)]-E-propenoate(10.9 g, 88%) as a white foam: R_(f)=0.60 (50% EtOAc in hexanes); IR(cm⁻¹) 3321, 1710; ¹H NGR (CDCl₃) δ 1.27 (t, 3H, J=7.2), 1.42 (s, 9H),1.70-1.78 (m, 1H), 1.80-1.96 (m, 1H), 2.35 (t, 2H, J=7.0), 4.18 (q, 2H,J=7.2), 4.29 (s, br, 1H), 4.82-4.84 (m, 1H), 5.88 (dd, 1H, J=15.7, 1.6),6.79 (dd, 1H, J=15.7, 5.3), 6.92 (s, 1H), 7.19-7.34 (m, 15H); Anal.(C₃₃H₃₈N₂O₅) C, H, N.

[0198] Preparation of IntermediateEthyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-Propenoate.

[0199] HCl (3 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of ethyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (0.091 g, 0.17 mmol, 1equiv) in 1,4-dioxane (3 mL) at 23° C. The reaction mixture was stirredat 23° C. for 1.5 hours, then was concentrated under reduced pressure toafford crude ethyl-3-[H₂N-L-(Tr-Gln)]-E-propenoate•HCl as a viscous oil.This material was dissolved in CH₂Cl₂ (6 mL) andCBZ-L-LeuΨ[COCH₂]-D/L-Phe-OH (0.068 g, 0.17 mmol, 1.0 equiv) [preparedas described in: Harbeson, S. L., Rich, D. H., J. Med. Chem. 1989, 32,1378], 1-hydroxybenzotriazole hydrate (0.030 g, 0.22 mmol, 1.3 equiv),4-methylmorpholine (0.055 mL, 0.50 mmol, 3.0 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.042 g,0.22 mmol, 1.3 equiv) were added sequentially. The reaction mixture wasstirred for 21 hours at 23° C. and then partitioned between water (100mL) and EtOAc (2×100 mL). The combined organic layers were dried overNa₂SO₄ and were concentrated. Purification of the residue by flashcolumn chromatography (50% EtOAc in hexanes) providedethyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-propenoate (0.040 g,28%) as a white foam: R_(f)=0.44 (50% EtOAc in hexanes); IR (cm⁻¹) 3317,1712, 1667; ¹H NGR (CDCl₃, approximately 1:1 mixture of diastereomers) δ0.84-0.91 (m), 1.20-1.31 (m), 1.51-1.58 (m), 1.73-1.96 (m), 2.29-2.39(m), 2.51-2.72 (m), 2.94-3.07 (m), 4.11-4.30 (m), 4.47-4.50 (m), 4.84(d, J=7.8 Hz), 4.94-5.08 (m), 5.09 (s), 5.30 (d, J=7.2 Hz), 5.48 (d,J=14.3 Hz), 5.72-5.95 (m), 6.55-7.01 (m), 7.14-7.61 (m), 8.02-8.05 (m);Anal. (C₅₂H₅₇N₃O₇.0.75H₂O) C, H, N.

[0200] Preparation of ProductEthyl-3-(CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-Gln)-E-Propenoate.

[0201] Triisopropylsilane (0.10 mL) and trifluoroacetic acid (3 mL) wereadded sequentially to a solution ofethyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-propenoate (0.035 g,0.042 mmol) in CH₂Cl₂ (4 mL) producing a bright yellow solution. Thereaction mixture was stirred for 30 minutes at 23° C., then carbontetrachloride (4 mL) was added, and the mixture was concentrated underreduced pressure. The residue was purified by flash columnchromatography (5% CH₃OH in CH₂Cl₂) to affordethyl-3-(CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-Gln)-E-propenoate (0.014 g, 56%) asa white foam: R_(f)=0.39 (10% CH₃OH in CH₂Cl₂); IR (cm⁻¹) 3306, 1712,1661; ¹H NGR (CDCl₃, approximately 1:1 mixture of diastereomers) δ0.87-0.93 (m), 1.24-1.33 (m), 1.39-1.96 (m), 2.17-2.21 (m), 2.58-2.79(m), 2.87-3.09 (m), 4.10-4.27 (m), 4.44 (s, br), 4.55 (s, br), 5.01-5.10(m), 5.14-5.69 (m), 5.79 (s), 5.82-5.91 (m), 6.13 (d, J=7.5 Hz), 6.42(s, br), 6.59 (dd, J=16.0, 7.5 Hz), 6.74 (dd, J=15.6, 4.7 Hz), 7.16-7.39(m); Anal. (C₃₃H₄₃N₃O₇) C, H, N.

Example 2

[0202] Preparation of Compound 1:Ethyl-3-(CBZ-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate.

[0203] Preparation of Intermediate trans-7-Methyl-oct-4-enoic Acid.

[0204] A solution of isovaleraldehyde (8.61 g, 100 mmol, 1 equiv) in THF(30 mL) was added dropwise via addition funnel to a solution ofvinylmagnesium bromide (100 mL of a 1.0 M solution in THF, 100 mmol, 1.0equiv) in THF (150 mL) at 0° C. After the addition was completed, thereaction mixture was stirred for 30 minutes at 0° C., and then waspartitioned between saturated NH₄Cl (150 mL) and Et₂O (2×150 mL). Thecombined organic layers were dried over Na₂SO₄ and concentrated toafford a yellow oil. This material was combined (neat) with diethylmalonate (16.7 mL, 110 mmol, 1.1 equiv) and Ti(OEt)₄ (2.10 mL, 10.0mmol, 0.10 equiv), and the combination was heated to 160° C. for 1 hour(distilling out EtOH as it was formed). The reaction mixture was thenmaintained at 190° C. for 4 hours and then cooled to 60° C. EtOH (50 mL)and 6.0 M KOH (50 mL) were added sequentially, and the brown reactionmixture was refluxed for 4 hours. After cooling to 23° C., the reactionmixture was filtered through a medium frit, and the filtrate waspartitioned between water (150 mL) and Et₂O (2×150 mL). The aqueouslayer was then acidified to pH=2 (as indicated by pH paper) withconcentrated HCl and extracted with a 1:1 mixture of EtOAc and hexanes(2×150 mL). The combined organic layers were dried over Na₂SO₄,concentrated, and the residue was distilled at reduced pressure toafford trans-7-methyl-oct-4-enoic acid (4.62 g, 30%) as a colorlessliquid: bp: 115-120° C. (1 torr); ¹H NGR (CDCl₃) δ 0.86 (d, 6H, J=5.5),1.51-1.65 (m, 1H), 1.87 (t, 2H, J=6.5), 2.22-2.38 (m, 2H), 2.40-2.45 (m,2H), 5.34-5.52 (m, 2H); Anal. (C₉H₁₆O₂) C, H.

[0205] Preparation of Intermediate trans-7-Methyl-oct-4-enoic Acid(2R-Hydroxy-1R-methyl-2-phenyl-ethyl)-methyl Amide.

[0206] Oxalyl chloride (2.71 mL, 31.1 mmol, 1.05 equiv) was added to asolution of trans-7-methyl-oct-4-enoic acid (4.62 g, 29.6 mmol, 1 equiv)and N,N-dimethylformamide (0.03 mL, 0.39 mmol, 0.012 equiv) in benzene(100 mL) at 23° C. The reaction mixture was stirred at 23° C. for 2hours and then concentrated under reduced pressure. The resulting oilwas dissolved in THF (20 mL) and added via cannula to a solution of(1R,2R)-(−)-pseudoephedrine (4.45 g, 26.9 mmol, 0.91 equiv) andtriethylamine (4.50 mL, 32.3 mmol, 1.1 equiv) in THF (200 mL) at 0° C.The reaction mixture was stirred at 0° C. for 30 minutes and thenpartitioned between half-saturated NH₄Cl (150 mL) and EtOAc (2×150 mL).The combined organic layers were dried over Na₂SO₄, concentrated, andthe residue purified by flash column chromatography (gradient elution40→50% EtOAc in hexanes) to afford trans-7-methyl-oct-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (7.55 g, 93%) as aviscous oil: R_(f)=0.27 (50% EtOAc in hexanes); IR (cm⁻¹) 3382, 1622; ¹HNGR (CDCl₃, mixture of rotamers) δ 0.87 (d, J=6.5), 0.99 (d, J=6.8),1.11 (d, J=7.2), 1.53-1.66 (m), 1.86 (t, J=6.1), 2.26-2.54 (m), 2.82(s), 2.92 (s), 3.99-4.04 (m), 4.29 (s, br), 4.42-4.47 (m), 4.56-4.62(m), 5.37-5.51 (m), 7.26-7.36 (m); Anal. (C₁₉H₂₉NO₂) C, H, N.

[0207] Preparation of Intermediate trans-2S-Benzyl-7-methyl-oct-4-enoicAcid (2R-Hydroxy-1R-methyl-2-phenyl-ethyl)-methyl Amide.

[0208] n-Butyllithium (52.6 mL of a 1.6 M solution in hexanes, 84.2mmol, 2.1 equiv) was added to a suspension of anhydrous lithium chloride(11.9 g, 282 mmol, 7.0 equiv) and diisopropylamine (12.7 mL, 90.3 mmol,2.25 equiv) in THF (300 mL) at −78° C. The reaction mixture was stirredfor 20 minutes at −78° C. and then maintained at 0° C. for 5 minutes andsubsequently cooled again to −78° C. trans-7-Methyl-oct-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (12.2 g, 40.1 mmol, 1equiv) in THF (40 mL) was added via cannula, and the resulting solutionwas stirred at −78° C. for 1 hour, maintained at 0° C. for 15 minutes,stirred at 23° C. for 5 minutes, and then cooled again to 0° C. Benzylbromide (7.15 mL, 60.1 mmol, 1.5 equiv) was added, and the reactionmixture was stirred at 0° C. for 30 minutes and then partitioned betweenhalf-saturated NH₄Cl (200 mL) and a 1:1 mixture of EtOAc and hexanes(2×200 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated. Purification of the residue by flash column chromatography(gradient elution 20→40% EtOAc in hexanes) providedtrans-2S-benzyl-7-methyl-oct-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (12.0 g, 76%) as aviscous oil: R_(f)=0.54 (50% EtOAc in hexanes); IR (cm⁻¹) 3382, 1617; ¹HNGR (CDCl₃, mixture of rotamers) δ 0.81-0.90 (m), 1.42-1.61 (m),1.80-1.95 (m), 2.17-2.25 (m), 2.33-2.54 (m), 2.55 (s), 2.73-2.99 (m),3.05-3.16 (m), 3.93-4.00 (m), 4.31-4.51 (m), 5.25-5.56 (m), 7.14-7.37(m); Anal. (C₂₆H₃₅NO₂) C, H, N.

[0209] Preparation of Intermediate3R-Benzyl-5S-(1R-bromo-3-methyl-butyl)-dihydrofuran-2-one.

[0210] N-Bromosuccinimide (5.97 g, 33.5 mmol, 1.1 equiv) was added insmall portions over 5 minutes to a solution oftrans-2S-benzyl-7-methyl-oct-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (12.0 g, 30.5 mmol, 1equiv) and glacial acetic acid (8.73 mL, 152 mmol, 5.0 equiv) in a 4:1mixture of THF and H₂O (250 mL) at 0° C. The resulting yellow solutionwas stirred for 15 minutes at 0° C., then warmed to 23° C., andsubsequently refluxed for 1 hour. After cooling to 23° C., the reactionmixture was partitioned between half-saturated NaHCO₃ (300 mL) and a 1:1mixture of EtOAc and hexanes (2×200 mL). The combined organic layerswere dried over Na₂SO₄ and concentrated. Flash chromatographicpurification of the residue (5% EtOAc in hexanes) gave3R-benzyl-5S-(1R-bromo-3-methyl-butyl)-dihydrofuran-2-one (7.09 g, 65%)as a pale yellow oil: R_(f)=0.79 (30% EtOAc in hexanes); IR (cm⁻¹) 1777;¹H NGR (CDCl₃) δ 0.87 (d, 3H, J=6.5), 0.94 (d, 3H, J=6.9), 1.53-1.72 (m,2H), 1.82-1.93 (m, 1H), 2.10-2.20 (m, 1H), 2.23-2.33 (m, 1H), 2.83 (dd,1H, J=13.5, 8.9), 3.04-3.12 (m, 1H), 3.14-3.22 (m, 1H), 4.05-4.12 (m,1H), 4.23-4.29 (m, 1H), 7.20-7.36 (m, 5H); Anal. (C₁₆H₂₁BrO₂) C, H, N.

[0211] Preparation of Intermediate5S-(1S-Azido-3-methyl-butyl)-3R-benzyl-dihydrofuran-2-one.

[0212] A suspension of sodium azide (2.83 g, 43.5 mmol, 2.0 equiv) and3R-benzyl-5S-(1R-bromo-3-methyl-butyl)-dihydrofuran-2-one (7.09 g, 21.8mmol, 1 equiv) in N,N-dimethylformamide (50 mL) was heated at 50° C. for20 hours. The reaction mixture was cooled to 23° C. and partitionedbetween water (200 mL) and a 1:1 mixture of EtOAc and hexanes (2×200mL). The combined organic layers were dried over Na₂SO₄, concentrated,and the residue purified by flash column chromatography (10% EtOAc inhexanes) to give5S-(1S-azido-3-methyl-butyl)-3R-benzyl-dihydrofuran-2-one (3.26 g, 52%)as a colorless oil: R_(f)=0.47 (20% EtOAc in hexanes); IR (cm⁻¹) 2109,1775; ¹H NGR (CDCl₃) δ 0.93 (d, 3H, J=6.5), 0.94 (d, 3H, J=6.5),1.32-1.41 (m, 1H), 1.55-1.65 (m, 1H), 1.70-1.85 (m, 1H), 2.03-2.18 (m,2H), 2.80 (dd, 1H, J=13.5, 8.9), 3.05-3.22 (m, 2H), 3.27-3.33 (m, 1H),4.22-4.27 (m, 1H), 7.18-7.36 (m, 5H); Anal. (C₁₆H₂₁N₃O₂) C, H, N.

[0213] Preparation of Intermediate[1S-(4R-Benzyl-5-oxo-tetrahydrofuran-2S-yl)-3-methyl-butyl]-carbamicAcid tert-Butyl Ester.

[0214] A suspension of5S-(1S-azido-3-methyl-butyl)-3R-benzyl-dihydrofuran-2-one (3.26 g, 11.3mmol, 1 equiv) and Pd/C (10%, 0.40 g) in CH₃OH (60 mL) was stirred undera hydrogen atmosphere (balloon) for 2 hours. The reaction mixture wasfiltered through celite, concentrated, and the residue dissolved in1,4-dioxane (80 mL). N,N-diisopropylethylamine (3.94 mL, 22.6 mmol, 2.0equiv) and di-tert-butyl dicarbonate (3.70 g, 17.0 mmol, 1.5 equiv) wereadded sequentially, and the resulting solution was stirred at 23° C. for2 hours. The reaction mixture was then partitioned between water (150mL) and a 1:1 mixture of EtOAc and hexanes (2×150 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated. Purification ofthe residue by flash column chromatography (gradient elution, 10→15%EtOAc in hexanes) provided[1S-(4R-benzyl-5-oxo-tetrahydrofuran-2S-yl)-3-methyl-butyl]-carbamicacid tert-butyl ester (2.53 g, 62%) as a white solid: mp=84-86° C.;R_(f)=0.66 (30% EtOAc in hexanes); IR (cm⁻¹) 3338, 1767, 1704; ¹H NGR(CDCl₃) δ 0.89 (d, 3H, J=6.5), 0.90 (d, 3H, J=6.5), 1.18-1.32 (m, 1H),1.40 (s, 9H), 1.43-1.56 (m, 1H), 1.98-2.07 (m, 1H), 2.20-2.29 (m, 1H),2.78 (dd, 1H, J=13.7, 9.0), 2.91-3.01 (m, 1H), 3.15 (dd, 1H, J=13.7,4.4), 3.71-3.81 (m, 1H), 4.23-4.28 (m, 1H), 4.34 (d, 1H, J=9.7),7.16-7.33 (m, 6H); Anal. (C₂₁H₃₁NO₄) C, H, N.

[0215] Preparation of IntermediateEthyl-3-[BOC-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenoate.

[0216] Lithium hydroxide (7.2 mL of a 1 M aqueous solution, 7.2 mmol,5.0 equiv) was added to a solution of[1S-(4R-benzyl-5-oxo-tetrahydrofuran-2S-yl)-3-methyl-butyl]-carbamicacid tert-butyl ester (0.521 g, 1.44 mmol, 1 equiv) in DME (7 mL) at 23°C. The resulting suspension was stirred at 23° C. for 30 minutes andthen partitioned between 0.5 M HCl (100 mL) and EtOAc (2×100 mL). Thecombined organic layers were dried over Na₂SO₄, concentrated, and theresidue dissolved in CH₂Cl₂ (30 mL). 4-Methylmorpholine N-oxide (0.337g, 2.88 mmol, 2.0 equiv), powdered 4 Å molecular sieves (0.55 g), andtetrapropylammonium perruthenate (0.050 g, 0.142 mmol, 0.10 equiv) wereadded sequentially. The resulting dark reaction mixture was stirred for2.5 hours at 23° C., and then it was filtered through celite. Thefiltrate was concentrated under reduced pressure to provide a brown oilwhich was dissolved in CH₂Cl₂ (30 mL). Crudeethyl-3-[H₂N-L-(Tr-Gln)]-E-propenoate•HCl (2.87 mmol, 2.0 equiv,prepared as described in Example 1 for the preparation ofethyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-propenoate),1-hydroxybenzotriazole hydrate (0.409 g, 3.03 mmol, 2.1 equiv),4-methylmorpholine (0.633 mL, 5.76 mmol, 4.0 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.58 g,3.03 mmol, 2.1 equiv) were added sequentially, and the reaction mixturewas stirred for 15 hours at 23° C. and then partitioned between water(150 mL) and EtOAc (2×150 mL). The combined organic layers were driedover Na₂SO₄ and concentrated. Purification of the residue by flashcolumn chromatography (gradient elution 30→40% EtOAc in hexanes)provided ethyl-3-[BOC-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.502 g, 44%) as a white foam: R_(f)=0.49 (50% EtOAc in hexanes); IR(cm⁻¹) 3314, 1707, 1667; ¹H NGR (CDCl₃) δ 0.84-0.91 (m, 6H), 1.29 (t,3H, J=7.2); 1.39 (s, 9H), 1.42-1.61 (m, 4H), 1.98-2.05 (m, 1H), 2.35 (t,2H, J=7.2), 2.54 (d, 1H, J=16.2), 2.70 (dd, 1H, J=11.5, 5.6), 2.78-2.99(m, 3H), 4.07-4.10 (m, 1H), 4.17 (q, 2H, J=7.2), 4.47 (s, br, 1H), 4.58(d, 1H, J=7.5), 5.46 (d, 1H, J=15.8), 5.87 (d, 1H, J=8.7), 6.58 (dd, 1H,J=15.8, 5.1), 7.12-7.31 (m, 21H); Anal. (C₄₉H₅₉N₃O₇) C, H, N.

[0217] Preparation of IntermediateEthyl-3-[CBZ-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenoate.

[0218] HCl (5 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of ethyl-3-[BOC-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenoate(0.096 g, 0.120 mmol, 1 equiv) in 1,4-dioxane (5 mL). The reactionmixture was stirred at 23° C. for 30 minutes and then concentrated. Theresulting oil was dissolved in CH₂Cl₂ (10 mL), and 4-methylmorpholine(0.033 mL, 0.300 mmol, 2.5 equiv) and benzyl chloroformate (0.025 mL,0.175 mmol, 1.4 equiv) were added sequentially. The reaction mixture wasstirred for 30 minutes at 23° C. and then partitioned between water (100mL) and a 1:1 mixture of EtOAc and hexanes (2×100 mL). The combinedorganic layers were dried over Na₂SO₄, concentrated, and the residue waschromatographed on silica gel (40% EtOAc in hexanes) to affordethyl-3-[CBZ-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate (0.050 g, 50%)as a white foam: R_(f)=0.38 (50% EtOAc in hexanes); IR (cm⁻¹) 3318,1712, 1664; ¹H NGR (CDCl₃) δ 0.85-0.87 (m, 6H), 1.05-1.13 (m, 1H), 1.29(t, 3H, J=7.2); 1.35-1.42 (m, 2H), 1.97 (s, br, 1H), 2.29-2.31 (m, 2H),2.54 (d, 1H, J=16.5), 2.69-3.03 (m, 5H), 4.17 (q, 2H, J=7.2), 4.47 (s,br, 1H), 4.81 (d, 1H, J=7.5), 4.94-5.06 (m, 3H), 5.48 (d, 1H, J=15.6),5.87 (d, 1H, J=8.1), 6.58 (dd, 1H, J=15.6, 5.0), 7.12-7.32 (m, 26H);Anal. (C₅₂H₅₇N₃O₇.0.25 H₂O) C, H, N.

[0219] Preparation of ProductEthyl-3-(CBZ-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate.

[0220] Triisopropylsilane (0.10 mL) and trifluoroacetic acid (5 mL) wereadded sequentially to a solution ofethyl-3-[CBZ-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate (0.050 g,0.060 mmol) in CH₂Cl₂ (6 mL) producing a bright yellow solution. Thereaction mixture was stirred for 30 minutes at 23° C., then carbontetrachloride (4 mL) was added, and the mixture was concentrated underreduced pressure. The residue was purified by flash columnchromatography (5% CH₃OH in CH₂Cl₂) to affordethyl-3-(CBZ-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-propenoate (0.026 g, 73%) as awhite solid: mp=162-164° C.; R_(f)=0.66 (10% CH₃OH in CH₂Cl₂); IR (cm⁻¹)3412, 3292, 1718, 1689, 1650; ¹H NGR (CDCl₃) δ 0.92 (d, 6H, J=6.5), 1.30(t, 3H, J=7.2), 1.45-1.59 (m, 3H), 1.97-2.05 (m, 1H), 2.07 (d, 1H,J=15.9), 2.17 (s, br, 2H), 2.70-2.79 (m, 1H), 2.91-3.09 (m, 3H), 4.18(q, 2H, J=7.2), 4.23-4.27 (m, 1H), 4.54 (s, br, 1H), 5.03 (d, 1H,J=12.1), 5.08 (d, 1H, J=12.1), 5.23 (d, 1H, J=6.9), 5.38 (s, br, 1H),5.47 (d, 1H, J=15.6), 5.92 (d, 1H, J=8.7), 6.43 (s, br, 1H), 6.60 (dd,1H, J=15.6, 4.8), 7.16-7.39 (m, 11H); Anal. (C₃₃H₄₃N₃O₇) C, H, N.

Example 3

[0221] Preparation of Compound 2:Ethyl-3-(EtSCO-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate.

[0222] Preparation of IntermediateEthyl-3-[EtSCO-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenoate.

[0223] HCl (4 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of ethyl-3-[BOC-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.216 g, 0.269 mmol, 1 equiv) in 1,4-dioxane (6 mL). The reactionmixture was stirred at 23° C. for 1.5 hours and then concentrated. Theresulting oil was dissolved in CH₂Cl₂ (6 mL), cooled to 0° C., andN,N-diisopropylethylamine (0.094 mL, 0.540 mmol, 2.0 equiv) and ethylchlorothiolformate (0.034 mL, 0.326 mmol, 1.2 equiv) were addedsequentially. The reaction mixture was stirred for 1 hour at 0° C., andit then was partitioned between water (100 mL) and EtOAc (2×100 mL). Thecombined organic layers were dried over Na₂SO₄, concentrated, and theresidue was purified by flash column chromatography (40% EtOAc inhexanes) to affordethyl-3-[EtSCO-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate (0.130 g,61%) as a white foam: R_(f)=0.45 (50% EtOAc in hexanes); IR (cm⁻¹) 3307,1713, 1656; ¹H NGR (CDCl₃) δ 0.86 (d, 6H, J=6.5), 1.05-1.19 (m, 1H),1.21-1.39 (m, 8H), 1.41-1.58 (m, 2H), 1.96-2.05 (m, 1H), 2.28-2.35 (m,2H), 2.54 (d, 1H, J=14.6), 2.70 (dd, 1H, J=11.7, 5.8), 2.79-3.00 (m,4H), 4.17 (q, 2H, J=7.2), 4.41-4.45 (m, 2H), 5.40 (d, 1H, J=7.5), 5.49(dd, 1H, J=15.8, 1.6), 5.93 (d, 1H, J=8.4), 6.59 (dd, 1H, J=15.8, 5.1),7.10-7.31 (m, 21H); Anal. (C₄₇H₅₅N₃O₆S) C, H, N.

[0224] Preparation of ProductEthyl-3-(EtSCO-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate

[0225] Ethyl-3-[EtSCO-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.120 g, 0.152 mmol) was dissolved in CH₂Cl₂ (5 mL), andtriisopropylsilane (0.10 mL) and trifluoroacetic acid (5 mL) were addedsequentially producing a bright yellow solution. The reaction mixturewas stirred for 30 minutes at 23° C., then carbon tetrachloride (6 mL)was added, and the mixture was concentrated under reduced pressure. Theresidue was purified by flash column chromatography (5% CH₃OH in CH₂Cl₂)to afford ethyl-3-(EtSCO-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-propenoate (0.056g, 68%) as a beige foam: R_(f)=0.55 (10% CH₃OH in CH₂Cl₂); IR (cm⁻¹)3385, 2293, 3199, 1717, 1653; ¹H NGR (CDCl₃) δ 0.91 (d, 3H, J=6.2), 0.92(d, 3H, J=6.5), 1.21-1.38 (m, 8H), 1.46-1.65 (m, 2H), 1.83-2.05 (m, 1H),2.61 (d, 1H, J=14.9), 2.74-3.09 (m, 7H), 4.18 (q, 2H, J=7.2), 4.49-4.56(m, 2H), 5.49 (d, 1H, J=15.6), 5.59 (s, br, 1H), 6.05 (d, 1H, J=8.7),6.20 (d, 1H, J=6.9), 6.49 (s, br, 1H), 6.62 (dd, 1H, J=15.6, 4.8),7.16-7.32 (m, 6H); Anal. (C₂₈H₄₁N₃O₆S) C, H, N.

Example 4

[0226] Preparation of Compound 6:Ethyl-3-(BnSCO-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate.

[0227] Preparation of Intermediate Benzyl Chlorothiolformate

[0228] Triethylamine (7.12 mL, 51.1 mmol, 1.0 equiv) was added to a 0°C. solution of benzyl mercaptan (6.0 mL, 51.1 mmol, 1 equiv) andtriphosgene (5.76 g, 19.4 mmol, 0.38 equiv) in CH₂Cl₂ (100 mL). Thereaction mixture was warmed to 23° C., stirred for 2 hours, and thenconcentrated under reduced pressure. The resulting white suspension wasslurried with Et₂O (100 mL) and filtered through a medium frit. Thefiltrate was concentrated under reduced pressure, and the resultingliquid was distilled under vacuum to provide benzyl chlorothiolformate(6.95 g, 73%) as a colorless liquid: bp=95-100° C. (8 torr); IR (cm⁻¹)1755; ¹H NGR (CDCl₃) δ 4.19 (s, 2H), 7.30-7.34 (m, 5H).

[0229] Preparation of IntermediateEthyl-3-[BnSCO-L-Leu□[COCH2]-L-Phe-L-(Tr-Gln)]-E-Propenoate.

[0230] HCl (5 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of ethyl-3-[BOC-L-Leu□[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.190 g, 0.237 mmol, 1 equiv) in 1,4-dioxane (5 mL). The reactionmixture was stirred at 23 □C for 1.5 hours and then concentrated. Theresulting oil was dissolved in CH₂Cl₂ (6 mL), and 4-methylmorpholine(0.078 mL, 0.709 mmol, 3.0 equiv) and benzyl chlorothiolformate (0.050mL, 0.331 mmol, 1.4 equiv) were added sequentially. The reaction mixturewas stirred for 1.5 hours at 23° C., and it then was partitioned betweenwater (100 mL) and a 1:1 mixture of EtOAc and hexanes (2×100 mL). Thecombined organic layers were dried over Na₂SO₄, concentrated, and theresidue was chromatographed on silica gel (40% EtOAc in hexanes) toafford ethyl-3-[BnSCO-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.117 g, 58%) as a white foam: R_(f)=0.44 (50% EtOAc in hexanes); IR(cm⁻¹) 3312, 1714, 1656; ¹H NGR (CDCl₃) δ 0.82-0.89 (m, 6H), 1.28 (t,3H, J=7.2), 1.43-1.57 (m, 2H), 1.95-2.05 (m, 1H), 2.31-2.36 (m, 2H),2.53 (d, 1H, J=14.6), 2.56-2.70 (m, 1H), 2.72-3.04 (m, 5H), 4.02-4.21(m, 4H), 4.44 (s, br, 2H), 5.41 (d, 1H, J=7.5), 5.48 (dd, 1H, J=15.8,1.6), 5.88 (d, 1H, J=8.1), 6.58 (dd, 1H, J=15.8, 5.1), 7.08-7.31 (m,26H); Anal. (C₅₂H₅₇N₃O₆S) C, H, N.

[0231] Preparation of ProductEthyl-3-(BnSCO-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate

[0232] Ethyl-3-[BnSCO-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.117 g, 0.137 mmol) was dissolved in CH₂Cl₂ (5 mL), andtriisopropylsilane (0.10 mL) and trifluoroacetic acid (5 mL) were addedsequentially producing a bright yellow solution. The reaction mixturewas stirred for 20 minutes at 23° C., then carbon tetrachloride (5 mL)was added, and the mixture was concentrated under reduced pressure. Theresidue was purified by flash column chromatography (5% CH₃OH in CH₂Cl₂)to afford ethyl-3-(BnSCO-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-propenoate (0.068g, 81%) as a white foam: R_(f)=0.52 (10% CH₃OH in CH₂Cl₂); IR (cm⁻¹)3299, 1717, 1650; ¹H NGR (CDCl₃) δ 0.91 (d, 6H, J=6.2), 1.29 (t, 3H,J=7.2), 1.36-1.63 (m, 3H), 1.80 (s, br, 1H), 1.94-2.05 (m, 1H),2.16-2.18 (m, 2H), 2.60 (d, 1H, J=16.5), 2.69-2.78 (m, 1H), 2.87-3.13(m, 3H), 4.05-4.21 (m, 4H), 4.52 (s, br, 2H), 5.47 (d, 1H, J=15.6), 5.57(s, 1H), 6.06 (d, 1H, J=8.7), 6.32 (d, 1H, J=7.2), 6.40 (s, 1H), 6.60(dd, 1H, J=15.6, 4.5), 7.15-7.29 (m, 10H); Anal. (C₃₃H₄₃N₃O₆S) C, H, N.

Example 5

[0233] Preparation of Compound 8:Ethyl-3-(CyPentylSCO-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate.

[0234] Preparation of Intermediate Cyclopentyl Chlorothiolformate.

[0235] Triethylamine (10.4 mL, 74.6 mmol, 1.0 equiv) was added to a 0°C. solution of cyclopentyl mercaptan (8.0 mL, 74.8 mmol, 1 equiv) andtriphosgene (8.43 g, 28.4 mmol, 0.38 equiv) in CH₂Cl₂. The reactionmixture was warmed to 23° C., stirred for 2 hours, and then concentratedunder reduced pressure. The resulting white suspension was slurried withEt₂O (100 mL) and filtered through a medium frit. The filtrate wasconcentrated under reduced pressure, and the resulting liquid wasdistilled under vacuum to provide cyclopentyl chlorothiolformate (10.4g, 85%) as a colorless liquid: bp=70-74° C. (1 torr); IR (cm⁻¹) 1756,830; ¹H NGR (C₆D₆) δ 1.01-1.23 (m, 6H), 1.49-1.60 (m, 2H), 3.20-3.29 (m,1H).

[0236] Preparation of IntermediateEthyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenoate

[0237] HCl (5 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of ethyl-3-[BOC-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.205 g, 0.256 mmol, 1 equiv) in 1,4-dioxane (6 mL). The reactionmixture was stirred at 23° C. for 1.5 hours and then concentrated. Theresulting oil was dissolved in CH₂Cl₂ (8 mL), cooled to 0° C., and4-methylmorpholine (0.070 mL, 0.637 mmol, 2.5 equiv) and cyclopentylchlorothiolformate (0.063 mL, 0.383 mmol, 1.5 equiv) were addedsequentially. The reaction mixture was stirred for 30 minutes at 0° C.,and it then was partitioned between water (100 mL) and a 1:1 mixture ofEtOAc and hexanes (2×100 mL). The combined organic layers were driedover Na₂SO₄, concentrated, and the residue was chromatographed on silicagel (40% EtOAc in hexanes) to provideethyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate (0.113g, 53%) as a white foam: R_(f)=0.44 (50% EtOAc in hexanes); IR (cm⁻¹)3310, 1713, 1654; ¹H NGR (CDCl₃) δ 0.85 (d, 6H, J=6.5), 1.02-1.12 (m,1H), 1.29 (t, 3H, J=7.2), 1.42-1.68 (m, 7H), 1.98-2.08 (m, 4H),2.31-2.35 (m, 2H), 2.55 (d, 1H, J=14.3), 2.70 (dd, 1H, J=11.8, 5.9),2.79-3.09 (m, 4H), 3.55-3.66 (m, 1H), 4.17 (q, 2H, J=7.2), 4.38 (s, br,1H), 4.48-4.49 (m, 1H), 5.33 (d, 1H, J=7.5), 5.50 (dd, 1H, J=15.9, 1.6),5.94 (d, 1H, J=8.4), 6.60 (dd, 1H, J=15.9, 5.0), 7.10-7.31 (m, 21H);Anal. (C₅₀H₅₉N₃O₆S) C, H, N.

[0238] Preparation of ProductEthyl-3-(CyPentylSCO-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate

[0239] Ethyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.090 g, 0.108 mmol) was dissolved in CH₂Cl₂ (6 mL), andtriisopropylsilane (0.10 mL) and trifluoroacetic acid (5 mL) were addedsequentially producing a bright yellow solution. The reaction mixturewas stirred for 20 minutes at 23° C., then carbon tetrachloride (4 mL)was added, and the mixture was concentrated under reduced pressure. Theresidue was purified by flash column chromatography (5% CH₃OH in CH₂Cl₂)to provide ethyl-3-(CyPentylSCO-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-propenoate(0.050 g, 79%) as a white foam: R_(f)=0.58 (10% CH₃OH in CH₂Cl₂); IR(cm⁻¹) 3385, 3298, 3199, 1717, 1652; ¹H NGR (CDCl₃) δ 0.89-0.93 (m, 6H),1.31 (t, 3H, J=7.2), 1.40-1.69 (m, 8H), 1.86 (s, br, 1H), 2.04-2.09 (m,4H), 2.21 (s, br, 2H), 2.58 (d, 1H, J=9.3), 2.64-2.79 (m, 1H), 2.89-3.07(m, 3H), 3.07-3.69 (m, 1H), 4.18 (q, 2H, J=7.2), 4.47 (s, br, 1H), 4.56(s, br, 1H), 5.49 (d, 1H, J=15.2), 5.57 (s, br, 1H), 6.07 (d, 1H,J=8.7), 6.15 (d, 1H, J=6.9), 6.54 (s, br, 1H), 6.62 (dd, 1H, J=15.2,4.8), 7.16-7.33 (m, 5H); Anal. (C₃₁H₄₅N₃O₆S) C, H, N.

Example 6

[0240] Preparation of Compound 4:Ethyl-3-(EtSCO-L-ValΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate.

[0241] Preparation of Intermediate trans-6-Methyl-hept-4-enoic Acid

[0242] A solution of isobutyraldehyde (9.59 g, 133 mmol, 1 equiv) in THF(50 mL) was added dropwise via addition funnel to a solution ofvinylmagnesium bromide (133 mL of a 1.0 M solution in THF, 133 mmol, 1.0equiv) in THF (300 mL) at 0° C. Upon completion of the addition, thereaction mixture was stirred for 30 minutes at 0° C., then ethyl malonylchloride (17.0 mL, 133 mmol, 1.0 equiv) was added. After stirring for 1hour at 0° C., the reaction mixture was partitioned between saturatedNH₄Cl (150 mL) and a 1:1 mixture of EtOAc and hexanes (2×200 mL). Thecombined organic layers were dried over Na₂SO₄ and concentrated.Purification of the residue by filtration through silica gel (elutingwith 5% EtOAc in hexanes) afforded the intermediate malonate ester (11.5g, 40% yield). This material was not characterized, but was combined(neat) with Ti(OEt)₄ (1.13 mL, 5.39 mmol, 0.10 equiv), heated to 190° C.for 4 hours, and then cooled to 60° C. EtOH (50 mL) and 6.0 M KOH (50mL) were added sequentially, and the brown reaction mixture was refluxedfor 4 hours. After cooling to 23° C., the reaction mixture was filteredthrough a medium frit, and the filtrate was partitioned between water(150 mL) and Et₂O (2×150 mL). The aqueous layer was then acidified topH=2 (as indicated by pH paper) with concentrated HCl and extracted witha 1:1 mixture of EtOAc and hexanes (2×150 mL). The combined organiclayers were dried over Na₂SO₄, concentrated, and the residue wasdistilled at reduced pressure to afford trans-6-methyl-hept-4-enoic acid(3.58 g, 47%) as a colorless liquid: bp: 107-112° C. (1 torr); IR (cm⁻¹)2960, 1711; ¹H NGR (CDCl₃) δ 0.96 (d, 6H, J=6.5) 2,18-2.45 (m, 5H),5.31-5.50 (m, 2H); Anal. (C₈H₁₄O₂) C, H.

[0243] Preparation of Intermediate trans-6-Methyl-hept-4-enoic Acid(2R-Hydroxy-1R-methyl-2-phenyl-ethyl)-methyl Amide.

[0244] Oxalyl chloride (2.25 mL, 25.8 mmol, 1.05 equiv) was added to asolution of trans-6-methyl-hept-4-enoic acid (3.50 g, 24.6 mmol, 1equiv) and N,N-dimethylformamide (0.03 mL, 0.39 mmol, 0.016 equiv) inbenzene (60 mL) at 23° C. The reaction mixture was stirred at 23° C. for2 hours and then concentrated under reduced pressure. The resulting oilwas dissolved in THF (20 mL) and added via cannula to a solution of1R,2R-(−)-pseudoephedrine (3.87 g, 23.4 mmol, 1.0 equiv) andtriethylamine (3.92 mL, 28.1 mmol, 1.2 equiv) in THF (150 mL) at 0° C.The reaction mixture was stirred at 0° C. for 30 minutes and thenpartitioned between half-saturated NH₄Cl (150 mL) and EtOAc (2×150 mL).The combined organic layers were dried over Na₂SO₄, concentrated, andthe residue purified by flash column chromatography (gradient elution40→50% EtOAc in hexanes) to afford trans-6-methyl-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (6.31 g, 93%) as aviscous oil: R_(f)=0.35 (50% EtOAc in hexanes); IR (cm⁻¹) 3382, 1622; ¹HNGR (CDCl₃, mixture of rotamers) δ 0.96 (d, J=6.8), 0.97 (d, J=6.5),1.11 (d, J=6.9), 2.18-2.59 (m), 2.82 (s), 2.92 (s), 3.99-4.04 (m),4.32-4.42 (m), 4.44-4.49 (m), 4.55-4.62 (m), 5.32-5.49 (m), 7.24-7.42(m); Anal. (C₁₈H₂₇NO₂) C, H, N.

[0245] Preparation of Intermediate trans-2S-Benzyl-6-methyl-hept-4-enoicAcid (2R-Hydroxy-1R-methyl-2-phenyl-ethyl)-methyl Amide.

[0246] n-Butyllithium (28.6 mL of a 1.6 M solution in hexanes, 45.8mmol, 2.1 equiv) was added to a suspension of anhydrous lithium chloride(6.47 g, 153 mmol, 7.0 equiv) and diisopropylamine (6.88 mL, 49.1 mmol,2.25 equiv) in THF (250 mL) at −78° C. The reaction mixture was stirredfor 20 minutes at −78° C., and it then was maintained at 0° C. for 5minutes and subsequently cooled again to −78° C.trans-6-Methyl-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (6.31 g, 21.8 mmol, 1equiv) in THF (40 mL) was added via cannula, and the resulting solutionwas stirred at −78° C. for 1 hour, maintained at 0° C. for 15 minutes,stirred at 23° C. for 5 minutes, and then cooled again to 0° C. Benzylbromide (3.89 mL, 32.7 mmol, 1.5 equiv) was added, the reaction mixturewas stirred at 0° C. for 30 minutes, and it then was partitioned betweenhalf-saturated NH₄Cl (200 mL) and a 1:1 mixture of EtOAc and hexanes(2×200 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated. Purification of the residue by flash column chromatography(gradient elution 20→40% EtOAc in hexanes) providedtrans-2S-benzyl-6-methyl-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (7.91 g, 96%) as aviscous oil: R_(f)=0.52 (50% EtOAc in hexanes); IR (cm⁻¹) 3383, 1616; ¹HNGR (CDCl₃, mixture of rotamers) δ 0.82-0.91 (m), 0.96 (d, J=6.5),1.24-1.27 (m), 2.14-2.47 (m), 2.56 (s), 2.72-2.99 (m), 3.04-3.15 (m),3.93-4.00 (m), 4.31-4.51 (m), 5.21-5.39 (m), 5.42-5.55 (m), 7.14-7.37(m); Anal. (C₂₅H₃₃NO₂) C, H, N.

[0247] Preparation of Intermediate3R-Benzyl-5S-(1R-bromo-2-methyl-propyl)-dihydrofuran-2-one.

[0248] N-Bromosuccinimide (3.89 g, 21.9 mmol, 1.05 equiv) was added insmall portions over 5 minutes to a solution oftrans-2S-benzyl-6-methyl-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (7.90 g, 20.8 mmol, 1equiv) and glacial acetic acid (5.96 mL, 104 mmol, 5.0 equiv) in a 4:1mixture of THF and H₂O (250 mL) at 0° C. The resulting yellow solutionwas stirred for 15 minutes at 0° C., then was warmed to 23° C., andsubsequently was refluxed for 1 hour. After cooling to 23° C., thereaction mixture was partitioned between half-saturated NaHCO₃ (300 mL)and a 1:1 mixture of EtOAc and hexanes (2×200 mL). The combined organiclayers were dried over Na₂SO₄ and concentrated. Flash chromatographicpurification of the residue (5% EtOAc in hexanes) gave3R-benzyl-5S-(1R-bromo-2-methyl-propyl)-dihydrofuran-2-one (5.09 g, 79%)as a white solid. Minor impurities were removed by recrystallizationfrom hexanes (2.51 g recovery): mp=75-76° C.; R_(f)=0.64 (30% EtOAc inhexanes); IR (cm⁻¹) 1774; ¹H NGR (CDCl₃) δ 0.93 (d, 3H, J=6.6), 0.99 (d,3H, J=6.9), 2.05-2.18 (m, 1H), 2.20-2.33 (m, 2H), 2.83 (dd, 1H, J=13.6,8.7), 2.95-3.05 (m, 1H), 3.17 (dd, 1H, J=13.6, 4.5), 3.89 (dd, 1H,J=9.0, 3.4), 4.32-4.39 (m, 1H), 7.20-7.36 (m, 5H); Anal. (C₁₅H₁₉BrO₂) C,H.

[0249] Preparation of Intermediate5S-(1S-Azido-2-methyl-propyl)-3R-benzyl-dihydrofuran-2-one.

[0250] A solution of Aliquat-336 (0.163 g, 0.403 mmol, 0.05 equiv) and3R-benzyl-5S-(1R-bromo-2-methyl-propyl)-dihydrofuran-2-one (2.51 g, 8.06mmol, 1 equiv) in toluene (60 mL) was treated with a solution of sodiumazide (2.10 g, 32.3 mmol, 4.0 equiv) in H₂O (10 mL). The resultingbiphasic mixture was heated to 70° C. and maintained at that temperaturefor 48 hours. The reaction mixture was cooled to 23° C. and partitionedbetween water (150 mL) and a 1:1 mixture of EtOAc and hexanes (2×150mL). The combined organic layers were dried over Na₂SO₄, concentrated,and the residue purified by flash column chromatography (gradientelution, 5→10% EtOAc in hexanes) to give5S-(1S-azido-2-methyl-propyl)-3R-benzyl-dihydrofuran-2-one (1.0 g, 45%)as a viscous oil: R_(f)=0.41 (20% EtOAc in hexanes); IR (cm⁻¹) 2105,1772; ¹H NGR (CDCl₃) δ 0.86-1.04 (m, 7H), 1.95-2.17 (m, 2H), 2.83 (dd,1H, J=13.2, 8.3), 2.92 (dd, 1H, J=6.4, 4.5), 3.05-3.21 (m, 2H),4.33-4.38 (m, 1H), 7.19-7.35 (m, 5H); Anal. (C₁₅H₁₉N₃O₂) C, H, N.

[0251] Preparation of Intermediate[1S-(4R-Benzyl-5-oxo-tetrahydrofuran-2S-yl)-2-methyl-propyl]-carbamicAcid tert-Butyl Ester.

[0252] A suspension of5S-(1S-azido-2-methyl-propyl)-3R-benzyl-dihydrofuran-2-one (1.00 g, 3.66mmol, 1 equiv) and Pd/C (10%, 0.090 g) in CH₃OH (60 mL) was stirredunder a hydrogen atmosphere (balloon) for 1 hour. The reaction mixturewas filtered through celite, concentrated, and the residue dissolved in1,4-dioxane (50 mL). N,N-diisopropylethylamine (1.28 mL, 7.35 mmol, 2.0equiv) and di-tert-butyl dicarbonate (1.20 g, 5.50 mmol, 1.5 equiv) wereadded sequentially, and the resulting solution was stirred at 23° C. for18 hours. The reaction mixture was then partitioned between water (100mL) and a 1:1 mixture of EtOAc and hexanes (2×150 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated. Purification ofthe residue by flash column chromatography (gradient elution, 10→15%EtOAc in hexanes) provided[1S-(4R-benzyl-5-oxo-tetrahydrofuran-2S-yl)-2-methyl-propyl]-carbamicacid tert-butyl ester (0.496 g, 39%) as a colorless oil: R_(f)=0.44 (20%EtOAc in hexanes); IR (cm⁻¹) 3340, 1768, 1708; ¹H NGR (CDCl₃) δ 0.92 (d,6H, J=6.9), 1.41 (s, 9H), 1.70-1.80 (m, 1H), 1.98-2.07 (m, 1H),2.14-2.24 (m, 1H), 2.81 (dd, 1H, J=13.6, 8.9), 2.92-3.01 (m, 1H), 3.13(dd, 1H, J=13.6, 4.4), 3.31-3.38 (m, 1H), 4.39-4.47 (m, 2H), 7.17-7.33(m, 5H); Anal. (C₂₀H₂₉NO₄) C, H, N.

[0253] Preparation of IntermediateEthyl-3-[BOC-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenoate.

[0254] Lithium hydroxide (7.14 mL of a 1 M aqueous solution, 7.14 mmol,5.0 equiv) was added to a solution of[1S-(4R-benzyl-5-oxo-tetrahydrofuran-2S-yl)-2-methyl-propyl]-carbamicacid tert-butyl ester (0.496 g, 1.43 mmol, 1 equiv) in DME (7 mL) at 23°C. The resulting suspension was stirred at 23° C. for 30 minutes andthen partitioned between 0.5 M HCl (100 mL) and EtOAc (2×100 mL). Thecombined organic layers were dried over Na₂SO₄, concentrated, and theresidue dissolved in CH₂Cl₂ (20 mL). 4-Methylmorpholine N-oxide (0.334g, 2.85 mmol, 2.0 equiv), powdered 4 Å molecular sieves (0.51 g), andtetrapropylammonium perruthenate (0.050 g, 0.142 mmol, 0.10 equiv) wereadded sequentially. The resulting dark reaction mixture was stirred for2.5 hours at 23° C. and then filtered through celite. The filtrate wasconcentrated under reduced pressure to provide a brown oil which wasdissolved in CH₂Cl₂ (30 mL). Crudeethyl-3-[H₂N-L-(Tr-Gln)]-E-propenoate•HCl, prepared as described inExample 1 for the preparation ofethyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-propenoate (1.86 mmol,1.3 equiv), 1-hydroxybenzotriazole hydrate (0.289 g, 2.14 mmol, 1.5equiv), 4-methylmorpholine (0.629 mL, 5.72 mmol, 4.0 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.411 g,2.14 mmol, 1.5 equiv) were added sequentially, and the reaction mixturewas stirred for 8 hours at 23° C. then partitioned between water (150mL) and EtOAc (2×150 mL). The combined organic layers were dried overNa₂SO₄ and concentrated. Purification of the residue by flash columnchromatography (40% EtOAc in hexanes) providedethyl-3-[BOC-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate (0.497 g, 44%)as an off-white foam: R_(f)=0.40 (50% EtOAc in hexanes); IR (cm⁻¹) 3314,1709, 1663; ¹H NGR (CDCl₃) δ 0.65 (d, 3H, J=6.9), 0.91 (d, 1H, J=6.9),1.29 (t, 3H, J=7.2), 1.40 (s, 9H), 1.94-2.20 (m, 2H), 2.30-2.35 (m, 2H),2.51 (d, 1H, J=17.1), 2.68 (dd, 1H, J=11.7, 5.8), 2.74-2.90 (m, 3H),3.04 (dd, 1H, J=17.3, 9.8), 4.05-4.09 (m, 1H), 4.17 (q, 2H, J=7.2),4.38-4.45 (m, 1H), 4.83 (d, 1H, J=8.1), 5.45 (d, 1H, J=15.7), 5.87 (d,1H, J=8.1), 6.57 (dd, 1H, J=15.7, 5.0), 7.10-7.31 (m, 21H); Anal.(C₄₈H₅₇N₃O₇) C, H, N.

[0255] Preparation of IntermediateEthyl-3-[EtSCO-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenoate.

[0256] HCl (5 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of ethyl-3-[BOC-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.225 g, 0.286 mmol, 1 equiv) in 1,4-dioxane (6 mL). The reactionmixture was stirred at 23° C. for 1.5 hours and then concentrated. Theresulting oil was dissolved in CH₂Cl₂ (10 mL), andN,N-diisopropylethylamine (0.125 mL, 0.718 mmol, 2.5 equiv) and ethylchlorothiolformate (0.040 mL, 0.384 mmol, 1.3 equiv) were addedsequentially. The reaction mixture was stirred for 1 hour at 23° C. andthen partitioned between water (100 mL) and a 1:1 mixture of EtOAc andhexanes (2×100 mL). The combined organic layers were dried over Na₂SO₄,concentrated, and the residue was chromatographed on silica gel (40%EtOAc in hexanes) to affordethyl-3-[EtSCO-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate (0.148 g,66%) as a white foam: R_(f)=0.37 (50% EtOAc in hexanes); IR (cm⁻¹) 3314,1714, 1653; ¹H NGR (CDCl₃) δ 0.68 (d, 3H, J=6.9), 0.92 (d, 3H, J=6.5),1.21-1.32 (m, 7H), 1.92-2.03 (m, 2H), 2.32-2.36 (m, 2H), 2.51 (dd, 1H,J=17.4, 2.2), 2.67 (dd, 1H, J=11.8, 5.9), 2.73-2.90 (m, 4H), 3.05 (dd,1H, J=17.6, 9.8), 4.17 (q, 2H, J=7.2), 4.39-4.44 (m, 2H), 5.46 (dd, 1H,J=15.7, 1.7), 5.62 (d, 1H, J=8.1), 5.96 (d, 1H, J=8.4), 6.57 (dd, 1H,J=15.7, 5.1), 7.10-7.12 (m, 2H), 7.15-7.31 (m, 19H); Anal. (C₄₆H₅₃N₃O₆S)C, H, N.

[0257] Preparation of ProductEthyl-3-(EtSCO-L-ValΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate.

[0258] Ethyl-3-[EtSCO-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.148 g, 0.191 mmol) was dissolved in CH₂Cl₂ (5 mL), andtriisopropylsilane (0.10 mL) and trifluoroacetic acid (5 mL) were addedsequentially producing a bright yellow solution. The reaction mixturewas stirred for 20 minutes at 23° C., then carbon tetrachloride (6 mL)was added, and the mixture was concentrated under reduced pressure. Theresidue was purified by flash column chromatography (5% CH₃OH in CH₂Cl₂)to afford ethyl-3-(EtSCO-L-ValΨ[COCH₂]-L-Phe-L-Gln)-E-propenoate (0.078g, 77%) as a white solid: mp=205° C. (dec); R_(f)=0.45 (10% CH₃OH inCH₂Cl₂); IR (cm⁻¹) 3424, 3304, 1715, 1658, 1640, 1624; ¹H NGR (DMSO-d₆)δ 0.75 (d, 3H, J=6.5), 0.82 (d, 3H, J=6.9), 1.14 (t, 3H, J=7.2), 1.21(t, 3H, J=7.2), 1.56-1.71 (m, 2H), 1.99-2.11 (m, 2H), 2.44-2.59 (m, 4H),2.70-2.85 (m, 3H), 2.93-2.95 (m, 1H), 4.06-4.18 (m, 3H), 4.31 (s, br,1H), 5.53 (d, 1H, J=15.6), 6.63 (dd, 1H, J=15.6, 5.1), 6.75 (s, 1H),7.14-7.26 (m, 6H), 8.06 (d, 1H, J=8.1), 8.33 (d, 1H, J=8.1); Anal.(C₂₇H₃₉N₃O₆S) C, H, N.

Example 7

[0259] Preparation of Compound 5:Ethyl-3-(BnSCO-L-ValΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate

[0260] Preparation of IntermediateEthyl-3-[BnSCO-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenoate.

[0261] HCl (8 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of ethyl-3-[BOC-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.272 g, 0.345 mmol, 1 equiv) in 1,4-dioxane (10 mL). The reactionmixture was stirred at 23° C. for 1.5 hours and then concentrated. Theresulting oil was dissolved in CH₂Cl₂ (10 mL), and 4-methylmorpholine(0.095 mL, 0.864 mmol, 2.5 equiv) and benzyl chlorothiolformate (0.068mL, 0.450 mmol, 1.3 equiv) were added sequentially. The reaction mixturewas stirred for 1 hour at 23° C. and then partitioned between water (100mL) and a 1:1 mixture of EtOAc and hexanes (2×100 mL). The combinedorganic layers were dried over Na₂SO₄, concentrated, and the residue waschromatographed on silica gel (40% EtOAc in hexanes) to affordethyl-3-[BnSCO-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate (0.160 g,55%) as a white foam: R_(f)=0.41 (50% EtOAc in hexanes); IR (cm⁻¹) 3316,1716, 1658; ¹H NGR (CDCl₃) δ 0.66 (d, 3H, J=6.9), 0.92 (d, 3H, J=6.5),1.29 (t, 3H, J=7.2), 1.93-2.03 (m, 2H), 2.32-2.36 (m, 2H), 2.50 (dd, 1H,J=17.7, 2.5), 2.67 (dd, 1H, J=11.5, 6.2), 2.73-2.89 (m, 3H), 3.07 (dd,1H, J=17.7, 10.0), 4.09 (s, 2H), 4.16 (q, 2H, J=7.2), 4.38-4.42 (m, 2H),5.46 (dd, 1H, J=15.8, 1.6), 5.65 (d, 1H, J=8.4), 5.95 (d, 1H, J=8.1),6.57 (dd, 1H, J=15.8, 5.1), 7.03 (s, 1H), 7.09-7.12 (m, 2H), 7.15-7.31(m, 23H); Anal. (C₅₁H₅₅N₃O₆S) C, H, N.

[0262] Preparation of ProductEthyl-3-(BnSCO-L-ValΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate.

[0263] Ethyl-3-[BnSCO-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.160 g, 0.191 mmol) was dissolved in CH₂Cl₂ (5 mL), andtriisopropylsilane (0.10 mL) and trifluoroacetic acid (5 mL) were addedsequentially producing a bright yellow solution. The reaction mixturewas stirred for 20 minutes at 23° C., then carbon tetrachloride (5 mL)was added, and the mixture was concentrated under reduced pressure. Theresidue was purified by flash column chromatography (5% CH₃OH in CH₂Cl₂)to provide ethyl-3-(BnSCO-L-ValΨ[COCH₂]-L-Phe-L-Gln)-E-propenoate (0.097g, 85%) as a white solid: mp=185-189° C.; R_(f)=0.55 (10% CH₃OH inCH₂Cl₂); IR (cm⁻¹) 3392, 3287, 1710, 1643; ¹H NGR (DMSO-d₆) δ 0.74 (d,3H, J=6.9), 0.82 (d, 3H, J=6.5), 1.21 (t, 3H, J=7.2), 1.62-1.70 (m, 2H),1.98-2.12 (m, 3H), 2.44-2.59 (m, 2H), 2.71-2.96 (m, 3H), 4.03 (s, 2H),4.10 (q, 2H, J=7.2), 4.19-4.23 (m, 1H), 4.32 (s, br, 1H), 5.54 (d, 1H,J=15.6), 6.64 (dd, 1H, J=15.6, 5.3), 6.75 (s, 1H), 7.14-7.28 (m, 11H),8.06 (d, 1H, J=8.1), 8.42 (d, 1H, J=7.8); Anal. (C₃₂H₄₁N₃O₆S) C, H, N.

Example 8

[0264] Preparation of Compound 7:Ethyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate.

[0265] Preparation of IntermediateEthyl-3-[CyPentylSCO-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenoate

[0266] HCl (6 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of ethyl-3-[BOC-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.248 g, 0.315 mmol, 1 equiv) in 1,4-dioxane (6 mL). The reactionmixture was stirred at 23° C. for 1.5 hours and then concentrated. Theresulting oil was dissolved in CH₂Cl₂ (15 mL), cooled to 0° C., and4-methylmorpholine (0.086 mL, 0.782 mmol, 2.5 equiv) and cyclopentylchlorothiolformate (0.067 mL, 0.407 mmol, 1.3 equiv) were addedsequentially. The reaction mixture was stirred for 30 minutes at 0° C.and then partitioned between water (100 mL) and a 1:1 mixture of EtOAcand hexanes (2×100 mL). The combined organic layers were dried overNa₂SO₄, concentrated, and the residue was chromatographed on silica gel(40% EtOAc in hexanes) to provideethyl-3-[CyPentylSCO-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate (0.126g, 49%) as a white foam: R_(f)=0.42 (50% EtOAc in hexanes); IR (cm⁻¹)3314, 1711, 1654; ¹H NGR (CDCl₃) δ 0.67 (d, 3H, J=6.9), 0.92 (d, 3H,J=6.9), 1.29 (t, 3H, J=7.2), 1.45-1.68 (m, 6H), 1.91-2.05 (m, 4H),2.31-2.34 (m, 2H), 2.51 (d, 1H, J=17.4), 2.67 (dd, 1H, J=11.5, 6.2),2.75-2.90 (m, 3H), 3.03 (dd, 1H, J=17.3, 9.8), 3.60-3.64 (m, 1H), 4.17(q, 2H, J=7.2), 4.36-4.44 (m, 2H), 5.46 (dd, 1H, J=15.8, 1.7), 5.57 (d,1H, J=8.1), 5.95 (d, 1H, J=8.4), 6.58 (dd, 1H, J=15.8, 5.0), 7.10-7.12(m, 2H), 7.19-7.31 (m, 19H); Anal. (C₄₉H₅₇N₃O₆S) C, H, N.

[0267] Preparation of ProductEthyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate

[0268] Ethyl-3-[CyPentylSCO-L-ValΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.126 g, 0.154 mmol) was dissolved in CH₂Cl₂ (6 mL), andtriisopropylsilane (0.10 mL) and trifluoroacetic acid (6 mL) were addedsequentially producing a bright yellow solution. The reaction mixturewas stirred for 20 minutes at 23° C., then carbon tetrachloride (6 mL)was added, and the mixture was concentrated under reduced pressure. Theresidue was purified by flash column chromatography (5% CH₃OH in CH₂Cl₂)to afford ethyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-Phe-L-Gln)-E-propenoate(0.046 g, 52%) as a white solid: mp=200-204° C.; R_(f)=0.38 (10% CH₃OHin CH₂Cl₂); IR (cm⁻¹) 3431, 3261, 1717, 1642; ¹H NGR (DMSO-d₆) δ 0.75(d, 3H, J=6.5), 0.82 (d, 3H, J=6.5), 1.22 (t, 3H, J=7.2), 1.40-1.74 (m,7H), 1.95-2.10 (m, 6H), 2.43-2.59 (m, 2H), 2.71-2.84 (m, 2H), 2.93-2.95(m, 1H), 3.47-3.56 (m, 1H), 4.06-4.16 (m, 3H), 4.32 (s, br, 1H), 5.54(d, 1H, J=15.9), 6.64 (dd, 1H, J=15.9, 5.3), 6.75 (s, 1H), 7.15-7.27 (m,6H), 8.06 (d, 1H, J=7.8), 8.27 (d, 1H, J=8.1); Anal. (C₃₀H₄₃N₃O₆S) C, H,N.

Example 9

[0269] Preparation of Compound 3:(2,3-Dihydroindole)-3-(BnSCO-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-Propenamide.

[0270] Preparation of Intermediate[2-(2,3-Dihydro-indol-1-yl)-2-oxo-ethyl]-phosphonic Acid Diethyl Ester.

[0271] Oxalyl chloride (5.96 mL, 68.3 mmol, 1.05 equiv) was added to asolution of diethylphosphonoacetic acid (12.8 g, 65.0 mmol, 1 equiv) andN,N-dimethylformamide (0.03 mL, 0.39 mmol, 0.006 equiv) in benzene (150mL) at 23° C. The reaction mixture was stirred at 23° C. for 1 hour andthen concentrated under reduced pressure. The resulting oil wasdissolved in THF (30 mL) and added via cannula to a solution of indoline(7.38 g, 61.9 mmol, 0.95 equiv) and triethylamine (10.9 mL, 78.0 mmol,1.2 equiv) in THF (200 mL) at 0° C. The reaction mixture was stirred at0° C. for 15 minutes, and it then was partitioned between 0.5 M HCl (150mL) and EtOAc (2×150 mL). The combined organic layers were dried overNa₂SO₄ and concentrated to afford a tan solid. Recrystallization fromEt₂O provided [2-(2,3-dihydro-indol-1-yl)-2-oxo-ethyl]-phosphonic aciddiethyl ester (12.2 g, 63%) as a light brown solid: mp: 97-99° C.;R_(f)=0.06 (50% EtOAc in hexanes); IR (cm⁻¹) 3460, 1657, 1597, 1482; ¹HNGR (CDCl₃) δ 1.35 (t, 6H, J=7.2), 3.14 (d, 2H, J=22.4), 3.22 (d, 2H,J=8.4), 4.15-4.30 (m, 6H), 7.04 (t, 1H, J=7.0), 7.17-7.28 (m, 2H), 8.21(d, 1H, J=9.0); Anal. (C₁₄H₂₀NO₄P) C, H, N.

[0272] Preparation of Intermediate(2,3-Dihydroindole)-3-[BOC-L-(Tr-Gln)]-E-Propenamide.

[0273] Sodium bis(trimethylsilyl)amide (11.9 mL of a 1.0 M solution inTHF, 11.9 mmol, 1.0 equiv) was added to a solution of[2-(2,3-dihydro-indol-1-yl)-2-oxo-ethyl]-phosphonic acid diethyl ester(3.54 g, 11.9 mmol, 1.0 equiv) in THF (150 mL) at −78° C., and theresulting solution was stirred for 20 minutes at that temperature. Crude[BOC-L-(Tr-Gln)]-H (5.63 g, 11.9 mmol, 1 equiv), prepared as describedin Example 1, in THF (40 mL) was added via cannula, and the reactionmixture was stirred for 1 hour at −78° C., warmed to 0° C. for 10minutes, and partitioned between 0.5 M HCl (150 mL) and EtOAc (2×150mL). The organic layers were dried over Na₂SO₄ and concentrated.Purification of the residue by flash column chromatography (50% EtOAc inhexanes) provided (2,3-dihydroindole)-3-[BOC-L-(Tr-Gln)]-E-propenamide(6.35 g, 87%) as an off-white foam: R_(f)=0.28 (50% EtOAc in hexanes);IR (cm⁻¹) 3401, 3307, 1690, 1665; ¹H NGR (CDCl₃) δ 1.44 (s, 9H),1.76-2.05 (m, 2H), 2.37-4.06 (m, 2H), 3.11-3.22 (m, 2H), 4.02-4.16 (m,2H), 4.27-4.40 (m, 1H), 4.91-4.97 (m, 1H), 6.29 (d, 1H, J=14.9),6.77-6.96 (m, 2H), 6.98-7.05 (m, 1H), 7.14-7.37 (m, 17H), 8.25 (d, 1H,J=7.5); Anal. (C₃₉H₄₁N₃O₄) C, H, N.

[0274] Preparation of Intermediate(2,3-Dihydroindole)-3-[BOC-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenamide.

[0275] Lithium hydroxide (8.0 mL of a 1 M aqueous solution, 8.0 mmol,5.0 equiv) was added to a solution of[1S-(4R-benzyl-5-oxo-tetrahydrofuran-2S-yl)-3-methyl-butyl]-carbamicacid tert-butyl ester (0.576 g, 1.59 mmol, 1 equiv), prepared asdescribed in Example 1, in DME (8 mL) at 23° C. The resulting suspensionwas stirred at 23° C. for 45 minutes, and it then was partitionedbetween 0.5 M HCl (100 mL) and EtOAc (2×100 mL). The combined organiclayers were dried over Na₂SO₄, concentrated, and the residue dissolvedin CH₂Cl₂ (30 mL). 4-Methylmorpholine N-oxide (0.373 g, 3.18 mmol, 2.0equiv), powdered 4 Å molecular sieves (0.60 g), and tetrapropylammoniumperruthenate (0.056 g, 0.159 mmol, 0.10 equiv) were added sequentially.The resulting dark reaction mixture was stirred for 1 hour at 23° C. andthen filtered through celite. The filtrate was concentrated underreduced pressure to provide a brown oil which was dissolved in CH₂Cl₂(20 mL). Crude (2,3-dihydroindole)-3-(H₂N-L-Tr-Gln)-E-propenamide•HCl(prepared from (2,3-dihydroindole)-3-[BOC-L-(Tr-Gln)]-E-propenamide in amanner analogous to the method described in Example 1 for thepreparation of ethyl-3-[H₂N-L-(Tr-Gln)]-E-propenoate•HCl (2.39 mmol, 1.5equiv), 1-hydroxybenzotriazole hydrate (0.322 g, 2.38 mmol, 1.5 equiv),4-methylmorpholine (0.699 mL, 6.36 mmol, 4.0 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.322 g,2.38 mmol, 1.5 equiv) were added sequentially, and the reaction mixturewas stirred for 16 hours at 23° C. and then partitioned between water(100 mL) and EtOAc (2×100 mL). The combined organic layers were driedover Na₂SO₄ and concentrated. Purification of the residue by flashcolumn chromatography (40% hexanes in EtOAc) provided(2,3-dihydroindole)-3-[BOC-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenamide(0.568 g, 40%) as tan foam: R_(f)=0.64 (10% CH₃OH in CH₂Cl₂); IR (cm⁻¹)3304, 1700, 1665; ¹H NGR (CDCl₃) δ 0.86 (d, 6H, J=6.5), 0.97-1.12 (m,1H), 1.39 (s, 9H), 1.45-1.64 (m, 2H), 2.39-2.43 (m, 2H), 2.51 (d, 1H,J=15.6), 2.67 (dd, 1H, J=12.3, 7.0), 2.79-2.96 (m, 3H), 3.16 (s, br,2H), 4.01-4.20 (m, 5H), 4.52 (s, br, 1H), 4.63 (d, 1H, J=7.8), 6.09 (d,1H, J=14.7), 6.17 (d, 1H, J=8.1), 6.65 (dd, 1H, J=14.7, 5.8), 6.99-7.04(m, 1H), 7.11-7.39 (m, 23H), 8.26 (d, 1H, J=7.5).

[0276] Preparation of Product(2,3-Dihydroindole)-3-[BnSCO-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenamide

[0277] HCl (6 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of(2,3-dihydroindole)-3-[BOC-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenamide(0.204 g, 0.233 mmol, 1 equiv) in 1,4-dioxane (8 mL). The reactionmixture was stirred at 23° C. for 2.5 hours and then concentrated. Theresulting oil was dissolved in CH₂Cl₂ (8 mL), andN,N-diisopropylethylamine (0.081 mL, 0.465 mmol, 2.0 equiv) and benzylchlorothiolformate (0.042 mL, 0.279 mmol, 1.2 equiv) were addedsequentially. The reaction mixture was stirred for 2 hours at 23° C. andthen partitioned between water (100 mL) and a 1:1 mixture of EtOAc andhexanes (2×100 mL). The combined organic layers were dried over Na₂SO₄and concentrated to provide crude(2,3-dihydroindole)-3-[BnSCO-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenamideas an off-white foam. This material was dissolved in CH₂Cl₂ (5 mL), andtriisopropylsilane (0.075 mL), and trifluoroacetic acid (5 mL) wereadded sequentially producing a bright yellow solution. The reactionmixture was stirred for 30 minutes at 23° C., then carbon tetrachloride(4 mL) was added, and the mixture was concentrated under reducedpressure. The residue was purified by flash column chromatography (5%CH₃OH in CH₂Cl₂) to afford(2,3-dihydroindole)-3-(BnSCO-L-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-propenamide(0.016 g, 10%) as a white foam: R_(f)=0.51 (10% CH₃OH in CH₂Cl₂); ¹H NGR(CDCl₃) δ 0.92 (d, 6H, J=6.2), 1.11-1.25 (m, 1H), 1.33-1.36 (m, 1H),1.49-2.17 (m, 4H), 2.25 (s, br, 2H), 2.58 (d, 1H, J=17.1), 2.70-2.72 (m,1H), 2.94-3.17 (m, 4H), 4.01-4.04 (m, 2H), 4.11 (s, 2H), 4.52-4.58 (m,2H), 5.47 (s, br, 1H), 6.13 (d, 1H, J=13.4), 6.29 (s, br, 1H), 6.41 (s,br, 1H), 6.66 (s, br, 1H), 7.00-7.05 (m, 1H), 7.17-7.29 (m, 13H), 8.25(d, 1H, J=7.2).

Example 10

[0278] Preparation of Compound 10:Ethyl-3-(EtSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-Propenoate.

[0279] Preparation of Intermediatetrans-6-Methyl-2S-(4-methyl-benzyl)-hept-4-enoic Acid(2R-Hydroxy-1R-methyl-2-phenyl-ethyl)-methyl Amide.

[0280] n-Butyllithium (33.7 mL of a 1.6 M solution in hexanes, 53.9mmol, 2.15 equiv) was added to a suspension of anhydrous lithiumchloride (7.47 g, 176 mmol, 7.0 equiv) and diisopropylamine (8.09 mL,57.7 mmol, 2.3 equiv) in THF (260 mL) at −78° C. The reaction mixturewas stirred for 30 minutes at −78° C., maintained at 0° C. for 5minutes, and subsequently cooled again to −78° C.trans-6-Methyl-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (7.26 g, 25.1 mmol, 1equiv) in THF (50 mL) was added via cannula, and the resulting solutionwas stirred at −78° C. for 1.75 hours, maintained at 0° C. for 20minutes, stirred at 23° C. for 5 minutes, and then cooled again to 0° C.A solution of 4-methylbenzyl bromide (6.96 g, 37.6 mmol, 1.5 equiv) inTHF (15 mL) was added, and the reaction mixture was stirred at 0° C. for30 minutes and then partitioned between half-saturated NH₄Cl (230 mL)and a 1:1 mixture of EtOAc and hexanes (200 mL, 2×150 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated. Purification ofthe residue by flash column chromatography (gradient elution 20→40%EtOAc in hexanes) providedtrans-6-methyl-2S-(4-methyl-benzyl)-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (9.33 g, 95%) as aviscous oil: R_(f)=0.44 (40% EtOAc in hexanes); IR (cm⁻¹) 3378, 1619; ¹HNGR (CDCl₃, mixture of rotamers) δ 0.84-0.97 (m), 2.12-2.27 (m), 2.29(s), 2.35-2.46 (m), 2.58 (s), 2.66-2.84 (m), 2.86-3.05 (m), 3.91-4.13(m), 4.30-4.44 (m), 4.45-4.53 (m), 5.17-5.54 (m), 7.04 (s), 7.12-7.33(m); Anal. (C₂₆H₃₅NO₂) C, H, N.

[0281] Preparation of Intermediate5S-(1R-Bromo-2-methyl-propyl)-3R-(4-methyl-benzyl)-dihydrofuran-2-one

[0282] N-Bromosuccinimide (4.37 g, 24.6 mmol, 1.05 equiv) was added insmall portions over 10 minutes to a solution oftrans-6-methyl-2S-(4-methyl-benzyl)-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (9.21 g, 23.4 mmol, 1equiv) and glacial acetic acid (6.70 mL, 117 mmol, 5.0 equiv) in a 4:1mixture of THF and H₂O (250 mL) at 0° C. The resulting yellow solutionwas stirred for 15 minutes at 0° C., then warmed to 23° C., andsubsequently refluxed for 45 minutes. After cooling to 23° C., thereaction mixture was partitioned between half-saturated NaHCO₃ (200 mL)and a 1:1 mixture of EtOAc and hexanes (2×200 mL, 100 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated. Flashchromatographic purification of the residue (gradient elution 4→6% EtOAcin hexanes) gave5S-(1R-bromo-2-methyl-propyl)-3R-(4-methyl-benzyl)-dihydrofuran-2-one(6.45 g, 85%) as a white solid (containing approximately 5-10%unidentified impurities by ¹H NGR): mp=94-100° C.; R_(f)=0.64 (25% EtOAcin hexanes); IR (cm⁻¹) 1772; ¹H NGR (CDCl₃, major isomer) 6 0.93 (d, 3H,J=6.5), 0.99 (d, 3H, J=6.5), 2.05-2.18 (m, 1H), 2.21-2.28 (m, 2H), 2.33(s, 3H), 2.79 (dd, 1H, J=13.5, 9.0), 2.92-3.03 (m, 1H), 3.12 (dd, 1H,J=13.5, 4.5), 3.89 (dd, 1H, J=9.0, 3.4), 4.32-4.41 (m, 1H), 7.06-7.16(m, 4H); Anal. (C₁₆H₂₁BrO₂) C, H.

[0283] Preparation of Intermediate5S-(1S-Azido-2-methyl-propyl)-3R-(4-methyl-benzyl)-dihydrofuran-2-one

[0284] A suspension of sodium azide (2.55 g, 39.2 mmol, 2.0 equiv) and5S-(1R-bromo-2-methyl-propyl)-3R-(4-methyl-benzyl)-dihydrofuran-2-one(6.37 g, 19.6 mmol, 1 equiv) in N,N-dimethylformamide (63 mL) was heatedat 50° C. for 48 hours. The reaction mixture was cooled to 23° C. andpartitioned between water (370 mL) and a 1:1 mixture of EtOAc andhexanes (2×370 mL, 200 mL). The combined organic layers were dried overNa₂SO₄, concentrated, and the residue purified by flash columnchromatography (gradient elution 8→12% EtOAc in hexanes) to give5S-(1S-azido-2-methyl-propyl)-3R-(4-methyl-benzyl)-dihydrofuran-2-one(3.33 g, 59%) as a colorless oil (containing approximately 5-10%unidentified impurities by ¹H NGR): R_(f)=0.52 (25% EtOAc in hexanes);IR (cm⁻¹) 2097, 1772; ¹H NGR (CDCl₃, major isomer) δ 0.97 (d, 3H,J=6.5), 1.01 (d, 3H, J=6.8), 1.94-2.06 (m, 1H), 2.07-2.13 (m, 2H), 2.33(s, 3H), 2.79 (dd, 1H, J=13.2, 8.2), 2.92 (dd, 1H, J=6.8, 4.4),3.02-3.17 (m, 2H), 4.32-4.40 (m, 1H), 7.07-7.15 (m, 4H); Anal.(C₁₆H₂₁N₃O₂) C, H, N.

[0285] Preparation of Intermediate{2-Methyl-1S-[4R-(4-methyl-benzyl)-5-oxo-tetrahydrofuran-2S-yl]-propyl}-carbamicAcid tert-Butyl Ester

[0286] A suspension of5S-(1S-azido-2-methyl-propyl)-3R-(4-methyl-benzyl)-dihydrofuran-2-one(3.24 g, 11.3 mmol, 1 equiv) and Pd/C (10%, 0.25 g) in CH₃OH (190 mL)was stirred under a hydrogen atmosphere (balloon) for 1 hour. Thereaction mixture was vacuum filtered through Whatman #3 paper,concentrated, and the residue dissolved in 1,4-dioxane (100 mL).N,N-diisopropylethylamine (3.93 mL, 22.6 mmol, 2.0 equiv) anddi-tert-butyl dicarbonate (3.69 g, 16.9 mmol, 1.5 equiv) were addedsequentially, and the resulting solution was stirred at 23° C. for 16hours. The reaction mixture was then partitioned between water (200 mL)and a 1:1 mixture of EtOAc and hexanes (2×300 mL, 200 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated. Purification ofthe residue by flash column chromatography (gradient elution, 11→17%EtOAc in hexanes) provided{2-methyl-1S-[4R-(4-methyl-benzyl)-5-oxo-tetrahydrofuran-2S-yl]-propyl}-carbamicacid tert-butyl ester (1.71 g, 42%) as a viscous oil: R_(f)=0.54 (25%EtOAc in hexanes); IR (cm⁻¹) 3331, 1766, 1696; ¹H NGR (CDCl₃) 8 0.92 (d,3H, J=6.5), 0.93 (d, 3H, J=6.5), 1.41 (s, 9H), 1.72-1.83 (m, 1H),1.96-2.06 (m, 1H), 2.11-2.22 (m, 1H), 2.31 (s, 3H), 2.77 (dd, 1H,J=13.5,8.7), 2.88-2.99 (m, 1H), 3.08 (dd, 1H, J=13.5, 4.5), 3.30-3.38 (m, 1H),4.38-4.44 (m, 1H), 4.49 (d, 1H, J=10.0), 7.04-7.13 (m, 4H); Anal.(C₂₁H₃₁NO₄) C, H, N.

[0287] Preparation of IntermediateEthyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-Propenoate

[0288] Lithium hydroxide (7.9 mL of a 1 M aqueous solution, 7.9 mmol,5.0 equiv) was added to a solution of{2-methyl-1S-[4R-(4-methyl-benzyl)-5-oxo-tetrahydrofuran-2S-yl]-propyl}-carbamicacid tert-butyl ester (0.570 g, 1.58 mmol, 1 equiv) in DME (15 mL) at23° C. The resulting suspension was stirred at 23° C. for 30 minutes andthen partitioned between 10% KHSO₄ (35 mL) and CH₂Cl₂ (100 mL, 2×70 mL).The combined organic layers were dried over Na₂SO₄, concentrated, andthe residue dissolved in CH₂Cl₂ (25 mL). Powdered 4 Å molecular sieves(0.56 g), 4-methylmorpholine N-oxide (0.369 g, 3.15 mmol, 2.0 equiv),and tetrapropylammonium perruthenate (0.055 g, 0.16 mmol, 0.10 equiv)were added sequentially. The resulting dark reaction mixture was stirredfor 1.33 hours at 23° C., and the mixture then was vacuum filteredthrough Whatman #3 paper and then through Whatman #5 paper. The filtratewas concentrated under reduced pressure to provide a dark residue whichwas dissolved in CH₂Cl₂ (25 mL). Crudeethyl-3-[H₂N-L-(Tr-Gln)]-E-propenoate•HCl (see preparation ofethyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-propenoate, 1.90 mmol,1.2 equiv), 4-methylmorpholine (0.693 mL, 6.30 mmol, 4.0 equiv),1-hydroxybenzotriazole hydrate (0.320 g, 2.37 mmol, 1.5 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.453 g,2.36 mmol, 1.5 equiv) were added sequentially, and the reaction mixturewas stirred for 20 hours at 23° C. and then partitioned between water(70 mL) and EtOAc (2×100 mL, 50 mL). The combined organic layers weredried over Na₂SO₄ and were concentrated. Purification of the residue byflash column chromatography (38% EtOAc in hexanes) providedethyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-CH₃) Phe-L-(Tr-Gln)]-E-propenoate (0.677g, 54%) as an off-white foam: R_(f)=0.44 (50% EtOAc in hexanes); IR(cm⁻¹) 3307, 1708, 1666; ¹H NGR (CDCl₃) δ 0.64 (d, 3H, J=6.8), 0.91 (d,3H, J=6.5), 1.28 (t, 3H, J=7.2), 1.40 (s, 9H), 1.55-1.67 (m, 1H),1.92-2.04 (m, 2H), 2.31 (s, 3H), 2.32-2.40 (m, 2H), 2.45-2.54 (m, 1H),2.63 (dd, 1H, J=11.5, 5.9), 2.68-2.87 (m, 2H), 3.03 (dd, 1H, J=17.4,10.0), 4.04-4.51 (m, 1H), 4.17 (q, 2H, J=7.2), 4.41-4.52 (m, 1H), 4.83(d, 1H, J=8.4), 5.49 (d, 1H, J=15.7), 5.86 (d, 1H, J=8.4), 6.60 (dd, 1H,J=15.7, 5.0), 7.00 (d, 2H, J=8.0), 7.09 (d, 2H, J=8.0); Anal.(C₄₉H₅₉N₃O₇) C, H, N.

[0289] Preparation of IntermediateEthyl-3-[EtSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-Propenoate

[0290] HCl (3 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution ofethyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gin)]-E-propenoate (0.315g, 0.393 mmol, 1 equiv) in 1,4-dioxane (3 mL). The reaction mixture wasstirred at 23° C. for 2 hours and then concentrated. The resulting oilwas dissolved in CH₂Cl₂ (5 mL), and N,N-diisopropylethylamine (0.205 mL,1.18 mmol, 3.0 equiv) and ethyl chlorothiolformate (0.061 mL, 0.59 mmol,1.5 equiv) were added sequentially. The reaction mixture was stirred for2 hours at 23° C. and then partitioned between brine (30 mL) and CH₂Cl₂(3×30 mL). The combined organic layers were dried over Na₂SO₄,concentrated, and the residue was chromatographed on silica gel (40%EtOAc in hexanes) to affordethyl-3-[EtSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate(0.220 g, 71%) as a white foam: R_(f)=0.29 (40% EtOAc in hexanes); IR(cm⁻¹) 3307, 1713, 1655; ¹H NGR (CDCl₃) δ 0.67 (d, 3H, J=6.8), 0.92 (d,3H, J=6.8), 1.25 (t, 3H, J =7.4), 1.28 (t, 3H, J=7.4), 1.55-1.67 (m,1H), 1.91-2.03 (m, 2H), 2.31 (s, 3H), 2.32-2.37 (m, 2H), 2.49 (dd, 1H,J=17.7, 2.2), 2.62 (dd, 1H, J=11.7, 6.1), 2.69-2.83 (m, 2H), 2.84 (q,2H, J=7.4), 3.04 (dd, 1H, J=17.7, 10.0), 4.17 (q, 2H, J=7.4), 4.35-4.51(m, 2H), 5.50 (dd, 1H, J=15.6, 1.6), 5.62 (d, 1H, J=8.1), 5.95 (d, 1H,J=8.1), 6.60 (dd, 1H, J=15.6, 5.1), 6.99 (d, 2H, J=8.1), 7.09 (d, 2H,J=8.1), 7.18-7.31 (m, 16H); Anal. (C₄₇H₅₅N₃O₆S) C, H, N.

[0291] Preparation of ProductEthyl-3-(EtSCO-L-ValΨ[COCH₂]-L-p-CH₃)Phe-L-Gln)-E-Propenoate

[0292] Ethyl-3-[EtSCO-L-ValΨ[COCH₂]-L-p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate (0.169 g, 0.214 mmol) was dissolved in CH₂Cl₂ (6 mL), andtriisopropylsilane (0.13 mL) and trifluoroacetic acid (3 mL) were addedsequentially producing a bright yellow solution. The reaction mixturewas stirred for 30 minutes at 23° C. and then concentrated under reducedpressure. The residue was purified by flash column chromatography (6%CH₃OH in CH₂Cl₂) to affordethyl-3-(EtSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-propenoate (0.072 g,63%) as a white solid: mp=220° C. (dec); R_(f)=0.11 (5% CH₃OH inCH₂Cl₂); IR (cm⁻¹) 3425, 3307, 1713, 1655; ¹H NGR (DMSO-d₆) δ 0.74 (d,3H, J=6.8), 0.82 (d, 3H, J=6.8), 1.14 (t, 3H, J=7.3), 1.20 (t, 3H,J=7.2), 1.54-1.76 (m, 2H), 1.98-2.13 (m, 3H), 2.23 (s, 3H), 2.40-2.54(m, 2H), 2.65-2.84 (m, 2H), 2.73 (q, 2H, J=7.3), 2.86-2.96 (m, 1H),4.04-4.19 (m, 3H), 4.26-4.37 (m, 1H), 5.55 (dd, 1H, J =15.7, 1.6), 6.65(dd, 1H, J=15.7, 5.3), 6.73 (s, 1H), 7.03 (s, 4H), 7.14 (s, 1H), 8.03(d, 1H, J=8.1), 8.31 (d, 1H, J=7.8); Anal. (C₂₈H₄₁N₃O₆S) C, H, N.

Example 11

[0293] Preparation of Compound 11:Ethyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-Propenoate

[0294] Preparation of IntermediateEthyl-3-[CyPentylSCO-L-ValY[COCH₂]-L-p-CH₃)Phe-L-(Tr-Gln)]-E-Propenoate

[0295] HCl (3 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution ofethyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate (0.304g, 0.379 mmol, 1 equiv) in 1,4-dioxane (3 mL). The reaction mixture wasstirred at 23° C. for 2.3 hours and then concentrated. The resulting oilwas dissolved in CH₂Cl₂ (5 mL), and N,N-diisopropylethylamine (0.198 mL,1.14 mmol, 3.0 equiv) and cyclopentyl chlorothiolformate (0.094 mL,0.571 mmol, 1.5 equiv) were added sequentially. The reaction mixture wasstirred for 3 hours at 23° C. and then partitioned between brine (30 mL)and CH₂Cl₂ (3×30 mL). The combined organic layers were dried overNa₂SO₄, concentrated, and the residue was chromatographed on silica gel(40% EtOAc in hexanes) to provideethyl-3-[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate(0.198 g, 63%) as a white foam: R_(f)=0.29 (40% EtOAc in hexanes); IR(cm⁻¹) 3307, 1713, 1655; ¹H NGR (CDCl₃) δ 0.67 (d, 3H, J=6.8), 0.92 (d,3H, J=6.8), 1.28 (t, 3H, J=7.2), 1.45-1.74 (m, 7H), 1.90-2.11 (m, 4H),2.31 (s, 3H), 2.32-2.38 (m, 2H), 2.50 (dd, 1H, J=17.7, 2.5), 2.62 (dd,1H, J=11.7, 5.8), 2.70-2.87 (m, 2H), 3.02 (dd, 1H, J=17.7, 9.8),3.57-3.67 (m, 1H), 4.17 (q, 2H, J=7.2), 4.22-4.31 (m, 2H), 5.50 (dd, 1H,J=15.8, 1.6), 5.58 (d, 1H, J=8.4), 5.93 (d, 1H, J=8.1), 6.60 (dd, 1H,J=15.8, 5.1), 6.99 (d, 2H, J=7.9), 7.09 (d, 2H, J=7.9), 7.18-7.31 (m,16H); Anal. (C₅₀H₅₉N₃O₆S) C, H, N.

[0296] Preparation of ProductEthyl-3-(CyPentylSCO-L-ValY[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-Propenoate

[0297] Ethyl-3-[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate (0.156 g, 0.188 mmol) was dissolved in CH₂Cl₂ (6 mL), andtriisopropylsilane (0.12 mL) and trifluoroacetic acid (3 mL) were addedsequentially producing a bright yellow solution. The reaction mixturewas stirred for 30 minutes at 23° C. and then concentrated under reducedpressure. Et₂O (5 mL) was added to the residue producing a whiteprecipitate. The precipitate was filtered, washed with Et₂O (3×3mL), anddried to provideethyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-propenoate(0.096 g, 87%) as a white solid: mp=208-210° C. (dec); R_(f)=0.43 (10%CH₃OH in CH₂Cl₂); IR (cm⁻¹) 3425, 3295, 1713, 1649; ¹H NGR (DMSO-d₆) δ0.74 (d, 3H, J=6.5), 0.81 (d, 3H, J=6.5), 1.20 (t, 3H, J=7.2), 1.37-1.76(m, 8H), 1.91-2.12 (m, 5H), 2.23 (s, 3H), 2.39-2.54 (m, 2H), 2.65-2.83(m, 2H), 2.86-2.96 (m, 1H), 3.46-3.57 (m, 1H), 4.04-4.17 (m, 3H),4.26-4.37 (m, 1H), 5.55 (dd, 1H, J=15.7, 1.4), 6.65 (dd, 1H, J=15.7,5.4), 6.74 (s, 1H), 7.03 (s, 4H), 7.15 (s, 1H), 8.03 (d, 1H, J=8.4),8.25 (d, 1H, J=8.1); Anal. (C₃₁H₄₅N₃O₆S) C, H, N.

Example 12

[0298] Preparation of Compound 12:Ethyl-3-(CyPentylSCO-L-PheΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-Propenoate.

[0299] Preparation of Intermediate(1R/S-Oxiranyl-2S-phenyl-ethyl)-carbamic Acid tert-Butyl Ester (1:1mixture of isomers).

[0300] DMSO (30 mL) was added to sodium hydride (0.900 g of a 60%dispersion in mineral oil, 22.5 mmol, 2.1 equiv), and the resultingsuspension was heated to 70° C. for 20 min. The clear solution thusobtained was cooled to 23° C., and THF (40 mL) was added. The reactionmixture was then cooled to 0° C., and a solution of trimethylsulfoniumiodide (4.59 g, 22.5 mmol, 2.1 equiv) in DMSO (30 mL) was added viacannula over 2 min. After stirring for 1 min at 0° C., a solution ofBOC-L-Phe-H (prepared according to Luly, J. R.; Dellaria, J. F.;Plattner, J. J.; Soderquist, J, L.; Yi, N. J. Org. Chem. 1987, 52, 1487)(2.5 g, 10.7 mmol, 1 equiv) in THF (20 mL) was added via cannula over 5min. The reaction mixture was stirred at 0° C. for 2 h and thenpartitioned between 0.5 M HCl (150 mL) and a 1:1 mixture of EtOAc andhexanes (2×150 mL). The organic layers were dried over Na₂SO₄ andconcentrated. Purification of the residue by flash column chromatography(gradient elution, 15→20% EtOAc in hexanes) afforded(1R/S-oxiranyl-2S-phenyl-ethyl)-carbamic acid tert-butyl ester (1:1mixture of isomers, 1.71 g, 60%) as a colorless oil: R_(f)=0.29 (isomer#1), 0.35 (isomer #2) (20% EtOAc in hexanes); IR (cm⁻¹) 3347, 2977,1700; ¹H NGR (CDCl₃) δ_(—)1.38 (s), 1.43 (s), 2.57-3.03 (m), 3.76 (s,br), 3.98 (s, br), 4.11 (s, br), 4.48 (s, br), 4.88 (s, br), 7.18-7.38(m); Anal. (C₁₅H₂₁NO₃) C, H, N.

[0301] Preparation of Intermediate 3-p-Tolyl-propionic Acidcis-1S-Amino-2R-indanol-acetonide Amide.

[0302] Oxalyl chloride (3.07 mL, 35.2 mmol, 1.05 equiv) was added to asolution of 3-p-tolyl-propionic acid (5.50 g, 33.5 mmol, 1 equiv) andN,N-dimethylformamide (0.03 mL, 0.39 mmol, 0.012 equiv) in benzene (150mL) at 23° C. The reaction mixture was stirred at 23° C. for 3 h andthen concentrated. The resulting oil was dissolved in THF (30 mL) andadded to a 0° C. solution of (1S,2R)-cis-1-amino-2-indanol (5.0 g, 33.5mmol, 1.0 equiv) and Et₃N (5.14 mL, 36.9 mmol, 1.1 equiv) in THF (250mL). After stirring for 30 min at 0° C., the reaction mixture waspartitioned between half-saturated NH₄Cl (150 mL) and EtOAc (2×150 mL).The organic layers were dried over Na₂SO₄ and concentrated to afford awhite solid. This material was dissolved in a 2:1 mixture of CH₂Cl₂ and2-methoxypropene (150 mL), and the resulting solution was treated withmethanesulfonic acid (1.0 mL). After stirring 1 h at 23° C., thereaction mixture was partitioned between half-saturated NaHCO₃ (150 mL),and CH₂Cl₂ (2×100 mL). The combined organic layers were washed with H₂O(100 mL), dried over MgSO₄, and gravity filtered. The filtrate wasconcentrated, and the residue was purified by flash columnchromatography (20% EtOAc in hexanes) to provide 3-p-tolyl-propionicacid cis-1S-amino-2R-indanol-acetonide amide (5.67 g, 51%) as a paleorange oil: R_(f)=0.63 (50% EtOAc in hexanes); IR (cm⁻¹) 2931, 1646; ¹HNGR (CDCl₃) δ_(—)1.35 (s, 3H), 1.61 (s, 3H), 2.33 (s, 3H), 2.90-2.95 (m,2H), 3.05-3.09 (m, 4H), 4.66-4.69 (m, 1H), 5.05 (d, 1H, J=4.7),7.10-7.27 (m, 8H); Anal. (C₂₂H₂₅NO₂) C, H, N.

[0303] Preparation of Intermediate{1S-[4R-(4-Methyl-benzyl)-5-oxo-tetrahydro-furan-2S-yl]-2-phenyl-ethyl}-carbamicAcid tert-Butyl Ester.

[0304] n-Butyllithium (8.12 mL of a 1.6 M solution in hexanes, 13.0mmol, 2.0 equiv) was added to a solution of(1R/S-oxiranyl-2S-phenyl-ethyl)-carbamic acid tert-butyl ester (1:1mixture of isomers, 1.71 g, 6.49 mmol, 1 equiv) and 3-p-tolyl-propionicacid cis-1S-amino-2R-indanol-acetonide amide (2.18 g, 6.50 mmol, 1equiv) in THF (100 mL) at −78° C. The reaction mixture was stirred for10 min at −78° C., maintained at 0° C. for 1.5 h, and then partitionedbetween 0.5 M HCl (150 mL) and a 1:1 mixture of EtOAc and hexanes (2×150mL). The organic layers were dried over Na₂SO₄ and concentrated. Flashchromatographic purification of the residue (gradient elution, 20→30%EtOAc in hexanes) gave the coupling product (1.02 g, single isomer, 26%)as an orange oil contaminated with several minor impurities. Thismaterial was dissolved in a 5:1 mixture of toluene and CH₂Cl₂ (60 mL)and was treated with p-toluenesulfonic acid monohydrate (0.324 g, 1.70mmol, 1.0 equiv) at 23° C. After stirring 12 h at 23° C., the reactionmixture was filtered through a medium frit, and the filtrate waspartitioned between half-saturated NaHCO₃ (150 mL) and a 1:1 mixture ofEtOAc and hexanes (2×100 mL). The organic layers were dried over Na₂SO₄and concentrated. The residue was purified by flash columnchromatography (15% EtOAc in hexanes) to provide{1S-[4R-(4-methyl-benzyl)-5-oxo-tetrahydro-furan-2S-yl]-2-phenyl-ethyl}-carbamicacid tert-butyl ester (0.26 g, 37%) as a white foam: R_(f)=0.60 (30%EtOAc in hexanes); IR (cm⁻¹) 3336, 1768, 1703; ¹H NGR (CDCl₃) δ_(—)1.36(s, 9H), 1.90-1.99 (m, 1H), 2.15-2.27 (m, 1H), 2.30 (s, 3H), 2.74 (dd,1H, J=13.5, 8.6), 2.82-2.88 (m, 2H), 2.91-3.00 (m, 1H), 3.06 (dd, 1H,J=13.5, 4.5), 3.88-3.97 (m, 1H), 4.18-4.23 (m, 1H), 4.51 (d, 1H, J=9.7),7.01-7.08 (m, 4H), 7.17-7.31 (m, 5H); Anal. (C₂₅H₃₁NO₄) C, H, N.

[0305] Preparation of IntermediateEthyl-3-[BOC-L-PheΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-Propenoate.

[0306] Lithium hydroxide (3.17 mL of a 1 M aqueous solution, 3.17 mmol,5 equiv) was added to a solution of{1S-[4R-(4-methyl-benzyl)-5-oxo-tetrahydro-furan-2S-yl]-2-phenyl-ethyl}-carbamicacid tert-butyl ester (0.260 g, 0.635 mmol, 1 equiv) in DME (4 mL) at23° C. The resulting suspension was stirred at 23° C. for 30 min andthen partitioned between 0.5 M HCl (100 mL) and EtOAc (2×100 mL). Thecombined organic layers were dried over Na₂SO₄, concentrated, and theresidue dissolved in a 1:1 mixture of CH₃CN and CH₂Cl₂ (60 mL).4-Methylmorpholine N-oxide (0.149 g, 1.27 mmol, 2 equiv), powdered 4 Åmolecular sieves (0.50 g), and tetrapropylammonium perruthenate (0.022g, 0.063 mmol, 0.1 equiv) were added sequentially. The resulting darkreaction mixture was stirred for 1 h at 23° C. and then filtered throughcelite. The filtrate was concentrated under reduced pressure to providea brown oil which was dissolved in CH₂Cl₂ (20 mL). Crudeethyl-3-[H₂N-L-(Tr-Gin)]-E-propenoate•HCl (see preparation ofethyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-propenoate, 0.762 mmol,1.2 equiv), 1-hydroxybenzotriazole hydrate (0.112 g, 0.829 mmol, 1.3equiv), 4-methylmorpholine (0.280 mL, 2.55 mmol, 4 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.158 g,0.824 mmol, 1.3 equiv) were added sequentially, and the reaction mixturewas stirred for 18 h at 23° C. and then partitioned between water (100mL) and a 1:1 mixture of EtOAc and hexanes (2×100 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated. Purification ofthe residue by flash column chromatography (40% EtOAc in hexanes)providedethyl-3-[BOC-L-PheΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate (0.190g, 35%) as a white foam: R_(f)=0.43 (50% EtOAc in hexanes); IR (cm⁻¹)3314, 1710, 1665; ¹H NGR (CDCl₃) δ 1.28 (t, 3H, J=7.2), 1.35 (s, 9H),1.63-1.65 (m, 1H), 1.97-2.05 (m, 1H), 2.33 (s, 3H), 2.36-2.38 (m, 2H),2.48 (d, 1H, J=15.9), 2.57-2.64 (m, 2H), 2.75-3.00 (m, 4H), 4.17 (q, 2H,J=7.2), 4.25-4.32 (m, 1H), 4.52 (s, br, 1H), 4.77 (d, 1H, J=6.9), 5.53(dd, 1H, J=15.9, 1.6), 5.94 (d, 1H, J=8.4), 6.63 (dd, 1H, J=15.9, 5.0),6.99-7.09 (m, 5H), 7.11-7.32 (m, 20H); Anal. (C₅₃H₅₉N₃O₇) C, H, N.

[0307] Preparation of IntermediateEthyl-3-[CyPentylSCO-L-PheΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-Propenoate.

[0308] HCl (5 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution ofethyl-3-[BOC-L-PheΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate (0.190g, 0.224 mmol, 1 equiv) in 1,4-dioxane (6 mL). The reaction mixture wasstirred at 23° C. for 1.5 h and then concentrated. The resulting oil wasdissolved in CH₂Cl₂ (10 mL), cooled to 0° C., and 4-methylmorpholine(0.075 mL, 0.682 mmol, 3.0 equiv) and cyclopentyl chlorothiolformate(0.055 mL, 0.334 mmol, 1.5 equiv) were added sequentially. The reactionmixture was stirred for 1 h at 0° C. and then partitioned between water(100 mL) and a 1:1 mixture of EtOAc and hexanes (2×100 mL). The combinedorganic layers were dried over Na₂SO₄, concentrated, and the residue waschromatographed on silica gel (40% EtOAc in hexanes) to affordethyl-3-[CyPentylSCO-L-PheΨ[COCH₂]-L-(-CH₃)Phe-L-(Tr-Gln)]-E-propenoate(0.102 g, 50%) as an off-white foam: R_(f)=0.49 (50% EtOAc in hexanes);IR (cm⁻¹) 3316, 1718, 1655; ¹H NGR (CDCl₃) δ 1.29 (t, 3H, J=7.2),1.45-1.67 (m, 7H), 1.98-2.05 (m, 2H), 2.33 (s, 3H), 2.42 (d, 1H,J=15.9), 2.55-2.98 (m, 6H), 3.52-3.63 (m, 1H), 4.17 (q, 2H, J=7.2),4.38-4.54 9 m, 2H), 5.52-5.58 (m, 2H), 6.03 (d, 1H, J=8.4), 6.64 (dd,1H, J=15.9, 5.0), 6.95-7.08 (m, 5H), 7.11-7.32 (m, 21H); Anal.(C₅₄H₅₉N₃O₆S) C, H, N.

[0309] Preparation of ProductEthyl-3-(CyPentylSCO-L-PheΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-Propenoate.

[0310] Triisopropylsilane (0.10 mL) and trifluoroacetic acid (5 mL) wereadded sequentially to a solution ofethyl-3-[CyPentylSCO-L-PheΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate(0.075 g, 0.085 mmol) in CH₂Cl₂ (6 mL) producing a bright yellowsolution. The reaction mixture was stirred for 15 min at 23° C., thencarbon tetrachloride (5 mL) was added, and the mixture was concentratedunder reduced pressure. The residue was purified by flash columnchromatography (5% CH₃OH in CH₂Cl₂) to affordethyl-3-(CyPentylSCO-L-PheΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-propenoate(0.047 g, 87%) as a white foam: R_(f)=0.62 (10% CH₃OH in CH₂Cl₂); IR(cm⁻¹) 3286, 1718, 1637; ¹H NGR (DMSO-d₆) δ 1.20 (t, 3H, J=6.8),1.33-1.70 (m, 10H), 1.91-2.07 (m, 4H), 2.24 (s, 3H), 2.38-3.05 (m, 5H),3.42-3.46 (m, 1H), 4.07-4.11 (m, 2H), 4.34 (s, br, 2H), 5.58 (d, 1H,J=15.7), 6.67 (dd, 1H, J=15.7, 4.4), 6.76 (s, br, 1H), 7.02-7.25 (m,10H), 8.07 (d, 1H, J=7.8), 8.44 (d, 1H, J=7.5); Anal. (C₃₅H₄₅N₃O₆S) C,H, N.

Example 13

[0311] Preparation of Compound 13:Ethyl-3-(EtSCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-Gln)-E-Propenoate.

[0312] Preparation of Intermediate trans-6-Methyl-2S-(4-fluorobenzyl)-hept-4-enoic Acid (2R-Hydroxy-1R-methyl-2-phenyl-ethyl)-methylAmide.

[0313] n-Butyllithium (32.5 mL of a 1.6 M solution in hexanes, 52.0mmol, 3.1 equiv) was added to a suspension of anhydrous lithium chloride(7.18 g, 169 mmol, 10 equiv) and diisopropylamine (7.80 mL, 55.7 mmol,3.3 equiv) in THF (250 mL) at −78° C. The reaction mixture was stirredfor 30 min at −78° C., then was maintained at 0° C. for 5 min, andsubsequently was cooled again to −78° C. trans-6-Methyl-hept-4-enoicacid (2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (4.91 g, 17.0mmol, 1 equiv) in THF (50 mL) was added via cannula, and the resultingsolution was stirred at −78° C. for 1.75 h, maintained at 0° C. for 20min, stirred at 23° C. for 5 min, and then cooled again to 0° C. Asolution of 4-fluorobenzyl bromide (6.34 mL, 50.9 mmol, 3 equiv) in THF(15 mL) was added, and the reaction mixture was stirred at 0° C. for 30min, and it then was partitioned between half-saturated NH₄Cl (230 mL)and a 1:1 mixture of EtOAc and hexanes (200 mL, 2×150 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated. Purification ofthe residue by flash column chromatography (gradient elution 20→40%EtOAc in hexanes) providedtrans-6-methyl-2S-(4-fluoro-benzyl)-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (6.33 g, 94%) as aviscous oil: R_(f)=0.38 (40% EtOAc in hexanes); IR (cm⁻¹) 3378, 1614; ¹HNGR (CDCl₃, mixture of rotamers) δ 0.85-0.95 (m), 0.96 (d, J=6.8),2.10-2.32 (m), 2.34-2.46 (m), 2.58 (s), 2.67-2.79 (m), 2.82-2.94 (m),3.00-3.18 (m), 3.94 (br), 4.37-4.52 (m), 5.24-5.42 (m), 5.44-5.56 (m),6.89-7.01 (m), 7.08-7.14 (m), 7.19-7.38 (m); Anal. (C₂₅H₃₂FNO₂) C, H, N.Preparation of Intermediate5S-(1R-Bromo-2-methyl-propyl)-3R-(4-fluoro-benzyl)-dihydrofuran-2-one.

[0314] N-Bromosuccinimide (2.93 g, 16.5 mmol, 1.05 equiv) was added insmall portions over 10 min to a solution oftrans-6-methyl-2S-(4-fluoro-benzyl)-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (6.24 g, 15.7 mmol, 1equiv) and glacial acetic acid (4.49 mL, 78.4 mmol, 5 equiv) in a 4:1mixture of THF and H₂O (165 mL) at 0° C. The resulting yellow solutionwas stirred for 15 min at 0° C., then was warmed to 23° C., andsubsequently was refluxed for 45 min. After cooling to 23° C., thereaction mixture was partitioned between half-saturated NaHCO₃ (200 mL)and a 1:1 mixture of EtOAc and hexanes (2×200 mL, 100 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated. Flashchromatographic purification of the residue (gradient elution 4→6→10%EtOAc in hexanes) gave5S-(1R-bromo-2-methyl-propyl)-3R-(4-fluoro-benzyl)-dihydrofuran-2-one(4.14 g, 80%) as a pale yellow oil (containing approximately 5-10%unidentified impurities by ¹H NGR): R_(f)=0.56 (25% EtOAc in hexanes);IR (cm⁻¹) 1772; ¹H NGR (CDCl₃, major isomer) δ 0.94 (d, 3H, J=6.5), 1.00(d, 3H, J=6.8), 2.05-2.35 (m, 3H), 2.83 (dd, 1H, J=13.6, 8.4), 2.92-3.03(m, 1H), 3.11 (dd, 1H, J=13.6, 4.7), 3.90 (dd, 1H, J=9.0, 3.7),4.33-4.40 (m, 1H), 6.98-7.06 (m, 2H), 7.14-7.20 (m, 2H); Anal.(C₁₅H₁₈BrFO₂) C, H.

[0315] Preparation of Intermediate5S-(1S-Azido-2-methyl-propyl)-3R-(4-fluoro-benzyl)-dihydrofuran-2-one.

[0316] A suspension of sodium azide (1.90 g, 29.2 mmol, 2.5 equiv) and5S-(1R-bromo-2-methyl-propyl)-3R-(4-fluoro-benzyl)-dihydrofuran-2-one(3.85 g, 11.7 mmol, 1 equiv) in N,N-dimethylformamide (40 mL) was heatedat 50° C. for 67 hours. The reaction mixture was cooled to 23° C. andpartitioned between half-saturated NaCl (200 mL) and a 1:1:1 mixture ofEtOAc, hexanes, and acetone (2×200 mL, 100 mL). The combined organiclayers were dried over Na₂SO₄, concentrated, and the residue purified byflash column chromatography (gradient elution 9→12→17% EtOAc in hexanes)to give5S-(1S-azido-2-methyl-propyl)-3R-(4-fluoro-benzyl)-dihydrofuran-2-one(2.10 g, 62%) as a white solid (containing approximately 5-10%unidentified impurities by ¹H NGR): mp 91-96° C.; R_(f)=0.44 (25% EtOAcin hexanes); IR (cm⁻¹) 2097, 1772; ¹H NGR (CDCl₃, major isomer) δ 0.99(d, 3H, J=6.5), 1.02 (d, 3H, J=6.8), 1.95-2.20 (m, 3H), 2.78-2.88 (m,1H), 2.94 (dd, 1H, J=7.0, 4.2), 3.03-3.17 (m, 2H), 4.37-4.43 (m, 1H),6.97-7.09 (m, 2H), 7.14-7.21 (m, 2H). Preparation of Intermediate{2-Methyl-1S-[4R-(4-fluoro-benzyl)-5-oxo-tetrahydrofuran-2S-yl]-propyl}-carbamicAcid tert-Butyl Ester.

[0317] A suspension of5S-(1S-azido-2-methyl-propyl)-3R-(4-fluoro-benzyl)-dihydrofuran-2-one(2.02 g, 6.93 mmol, 1 equiv), di-tert-butyl dicarbonate (2.12 g, 9.71mmol, 1.4 equiv) and Pd/C (10%, 0.20 g) in CH₃OH (100 mL) was stirredunder a hydrogen atmosphere (balloon) for 16 hours. The reaction mixturewas vacuum filtered through Whatman #3 paper and concentrated.Purification of the residue by flash column chromatography (15% EtOAc inhexanes) provided{2-methyl-1S-[4R-(4-fluoro-benzyl)-5-oxo-tetrahydrofuran-2S-yl]-propyl}-carbamicacid tert-butyl ester (1.58 g, 62%) as a white foam: R_(f)=0.80 (5% MeOHin CH₂Cl₂); IR (cm⁻¹) 3331, 1766, 1702; ¹H NGR (CDCl₃) δ 0.93 (d, 3H,J=6.8), 0.95 (d, 3H, J=6.5), 1.41 (s, 9H), 1.71-1.83 (m, 1H), 1.95-2.06(m, 1H), 2.16-2.27 (m, 1H), 2.80 (dd, 1H, J=13.5, 8.6), 2.88-2.99 (m,1H), 3.09 (dd, 1H, J=13.5, 4.4), 3.32-3.40 (m, 1H), 4.42-4.48 (m, 2H),6.95-7.03 (m, 2H), 7.11-7.18 (m, 2H); Anal. (C₂₀H₂₈FNO₄) C, H, N.

[0318] Preparation of IntermediateEthyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-Propenoate.

[0319] Lithium hydroxide (9.62 mL of a 1 M aqueous solution, 9.62 mmol,5 equiv) was added to a solution of{2-methyl-1S-[4R-(4-fluoro-benzyl)-5-oxo-tetrahydrofuran-2S-yl]-propyl}-carbamicacid tert-butyl ester (0.703 g, 1.92 mmol, 1 equiv) in DME (25 mL) at23° C. The resulting suspension was stirred at 23° C. for 30 min, and itthen was partitioned between 10% KHSO₄ (50 mL) and CH₂Cl₂ (3×100 mL).The combined organic layers were dried over Na₂SO₄, concentrated, andthe residue dissolved in CH₂Cl₂ (30 mL). Powdered 4 521 molecular sieves(0.70 g), 4-methylmorpholine N-oxide (0.451 g, 3.85 mmol, 2 equiv), andtetrapropylammonium perruthenate (0.068 g, 0.19 mmol, 0.10 equiv) wereadded sequentially. The resulting dark reaction mixture was stirred for1.33 hours at 23° C., and it then was vacuum filtered through Whatman #3paper and then through Whatman #5 paper. The filtrate was concentratedunder reduced pressure to provide a dark residue which was dissolved inCH₂Cl₂ (30 mL). Crude ethyl-3-[H₂N-L-(Tr-Gln)]-E-propenoate•HCl (2.30mmol, 1.2 equiv, prepared as described in Example 1 for the preparationof ethyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-propenoate),4-methylmorpholine (0.846 mL, 7.69 mmol, 4 equiv),1-hydroxybenzotriazole hydrate (0.390 g, 2.89 mmol, 1.5 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.553 g,2.88 mmol, 1.5 equiv) were added sequentially, and the reaction mixturewas stirred for 19 hours at 23° C. and then partitioned between brine(100 mL) and CH₂Cl₂ (3×100 mL). The combined organic layers were driedover Na₂SO₄ and concentrated. Purification of the residue by flashcolumn chromatography (gradient elution 35→40% EtOAc in hexanes)provided ethyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.820 g, 53%) as a tan foam: R_(f)=0.50 (50% EtOAc in hexanes); IR(cm⁻¹) 3307, 1708, 1666; ¹H NGR (CDCl₃) δ 0.67 (d, 3H, J=6.8), 0.92 (d,3H, J=6.8), 1.28 (t, 3H, J=7.2), 1.40 (s, 9H), 1.53-1.67 (m, 1H),1.91-2.04 (m, 2H), 2.32-2.41 (m, 2H), 2.46-2.55 (m, 1H), 2.63 (dd, 1H,J=12.1, 5.9), 2.69-2.80 (m, 1H), 2.83 (dd, 1H, J=12.1, 8.2), 3.03 (dd,1H, J=17.7, 10.0), 4.05-4.11 (m, 1H), 4.17 (q, 2H, J=7.2), 4.40-4.50 (m,1H), 4.84 (d, 1H, J=8.4), 5.38 (d, 1H, J=15.7), 6.01 (d, 1H, J=8.4),6.60 (dd, 1H, J=15.7, 5.0), 6.92-6.99 (m, 2H), 7.03-7.12 (m, 3H),7.17-7.30 (m, 15H); Anal. (C₄₈H₅₆FN₃O₇) C, H, N.

[0320] Preparation of IntermediateEthyl-3-[EtSCO-L-ValΨ[COCH₂]-L-p-F)Phe-L-(Tr-Gln)]-E-Propenoate.

[0321] HCl (3 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution ofethyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gin)]-E-propenoate (0.271 g,0.336 mmol) in 1,4-dioxane (3 mL). The reaction solution was stirred at23° C. for 2 hours and then was concentrated. The residue was dissolvedin dry CH₂Cl₂ (5 mL). N,N-diisopropylethylamine (0.176 mL, 1.01 mmol, 3equiv) and ethyl chlorothiolformate (0.060 mL, 0.58 mmol, 1.7 equiv)were added sequentially. The reaction solution was stirred 2 hours at23° C. and then was partitioned between brine (30 mL) and CH₂Cl₂ (3×30mL). The combined organic phases were dried over Na₂SO₄, concentrated,and the residue was chromatographed on silica gel (gradient elution40→50% EtOAc in hexanes) to affordethyl-3-[EtSCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate (0.183g, 69%) as a white foam: R_(f)=0.43 (50% EtOAc in hexanes); IR (cm⁻¹)3307, 1708, 1655; ¹H NGR (CDCl₃) δ 0.70 (d, 3H, J=6.8), 0.93 (d, 3H,J=6.5), 1.25 (t, 3H, J=7.2), 1.29 (t, 3H, J=7.2), 1.55-1.66 (m, 1H),1.91-2.05 (m, 2H), 2.30-2.39 (m, 2H), 2.51 (dd, 1H, J=17.4, 2.5), 2.63(dd, 1H, J=12.1, 5.9), 2.69-2.90 (m, 4H), 3.02 (dd, 1H, J=17.4, 10.0),4.17 (q, 2H, J=7.2), 4.35-4.49 (m, 2H), 5.39 (dd, 1H, J=15.6, 1.7), 5.66(d, 1H, J=8.1), 6.12 (d, 1H, J=8.1), 6.60 (dd, 1H, J=15.6, 5.0),6.91-6.99 (m, 2H), 7.02-7.10 (m, 2H), 7.17-7.32 (m, 16H); Anal.(C₄₆H₅₂FN₃O₆S) C, H, N.

[0322] Preparation of ProductEthyl-3-(EtSCO-L-ValΨ[COCH₂]-L-p-F)Phe-L-Gln)-E-Propenoate.

[0323] Ethyl-3-[EtSCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.149 g, 0.188 mmol, 1 equiv) was dissolved in dry CH₂Cl₂ (6 mL).Triisopropylsilane (0.115 mL, 0.561 mmol, 3 equiv) and trifluoroaceticacid (3 mL) were added sequentially producing a bright yellow solution.This solution was stirred for 30 minutes and then concentrated. Theresidue was stirred in Et₂O (6 mL), and the solid was collected byfiltration, washed with Et₂O (3 mL then 2 mL), and then dried undervacuum to giveethyl-3-(EtSCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-Gln)-E-propenoate (0.090 g,87%) as a white solid: mp=214° C. (dec); R_(f)=0.49 (10% CH₃OH inCH₂Cl₂); IR (cm⁻¹) 3425, 3284, 1713, 1643; ¹H NGR (DMSO-d₆) δ 0.76 (d,3H, J=6.5), 0.82 (d, 3H, J=6.8), 1.15 (t, 3H, J=7.2), 1.21 (t, 3H,J=7.2), 1.53-1.75 (m, 2H), 1.99-2.14 (m, 3H), 2.52-2.85 (m, 6H),2.88-2.99 (m, 1H), 4.09 (q, 2H, J=7.2), 4.13-4.20 (m, 1H), 4.25-4.36 (m,1H), 5.40 (dd, 1H, J=15.6, 1.4), 6.61 (dd, 1H, J=15.6, 5.3), 6.74 (s,1H), 6.99-7.24 (m, 5H), 8.01 (d, 1H, J=8.1), 8.34 (d, 1H, J=8.1); Anal.(C₂₇H₃₈FN₃O₆S) C, H, N.

Example 14

[0324] Preparation of Compound 14:Ethyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-p-F)Phe-L-Gln)-E-Propenoate.

[0325] Preparation of IntermediateEthyl-3-[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-Propenoate.

[0326] HCl (3 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution ofethyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate (0.273 g,0.339 mmol) in 1,4-dioxane (3 mL). The reaction solution was stirred at23° C. for 2 hours and then concentrated. The residue was dissolved indry CH₂Cl₂ (5 mL). N,N-diisopropylethylamine (0.177 mL, 1.02 mmol, 3equiv) and cyclopentyl chlorothiolformate (0.095 mL, 1.7 equiv) wereadded sequentially. The reaction solution was stirred 2 hours at 23° C.and then partitioned between brine (30 mL) and CH₂Cl₂ (3×30 mL). Thecombined organic phases were dried over Na₂SO₄, concentrated, and theresidue was chromatographed on silica gel (40% EtOAc in hexanes) toaffordethyl-3-[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.166 g, 59%) as a white foam: R_(f)=0.24 (40% EtOAc in hexanes); IR(cm⁻¹) 3307, 1713, 1654; ¹H NGR (CDCl₃) δ 0.69 (d, 3H, J=6.8), 0.93 (d,3H, J=6.8), 1.29 (t, 3H, J=7.2), 1.47-1.75 (m, 7H), 1.91-2.12 (m, 4H),2.30-2.41 (m, 2H), 2.51 (dd, 1H, J=17.2, 2.3), 2.63 (dd, 1H, J=12.3,5.9), 2.69-2.80 (m, 1H), 2.84 (dd, 1H, J=12.3, 8.4), 3.01 (dd, 1H,J=17.2, 10.0), 3.57-3.67 (m, 1H), 4.17 (q, 2H, J=7.2), 4.33-4.50 (m,2H), 5.39 (dd, 1H, J=15.7, 1.7), 5.61 (d, 1H, J=7.8), 6.11 (d, 1H,J=8.1), 6.60 (dd, 1H, J=15.7, 4.8), 6.92-7.00 (m, 2H), 7.03-7.12 (m,3H), 7.18-7.32 (m, 15H); Anal. (C₄₉H₅₆FN₃O₆S.0.5 H₂O) C, H,N.

[0327] Preparation of ProductEthyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-p-F)Phe-L-Gln)-E-Propenoate.

[0328]Ethyl-3-[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.132 g, 0.158 mmol, 1 equiv) was dissolved in dry CH₂Cl₂ (6 mL).Triisopropylsilane (0.097 mL, 0.47 mmol, 3 equiv) and trifluoroaceticacid (3 mL) were added sequentially producing a bright yellow solution.This solution was stirred for 30 minutes and then concentrated. Theresidue was stirred in Et₂O (6 mL), and the solid was collected byfiltration, washed with Et₂O (3 mL then 2 mL), and then dried undervacuum to giveethyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-Gln)-E-propenoate (0.077g, 82%) as a white solid: mp=215° C. (dec); R_(f)=0.45 (10% CH₃OH inCH₂Cl₂); IR (cm⁻¹) 3413, 3296, 1715, 1649; ¹H NGR (DMSO-d₆) δ 0.75 (d,3H, J=6.5), 0.82 (d, 3H, J=6.5), 1.21 (t, 3H, J=6.9), 1.36-1.75 (m, 8H),1.92-2.14 (m, 5H), 2.52-2.85 (m, 4H), 2.87-2.99 (m, 1H), 3.47-3.58 (m,1H), 4.06-4.18 (m, 1H), 4.09 (q, 2H, J=6.9), 4.25-4.36 (m, 1H), 5.41 (d,1H, J=15.6), 6.61 (dd, 1H, J=15.6, 5.1), 6.74 (s, 1H), 6.98-7.23 (m,5H), 8.01 (d, 1H, J=8.4), 8.28 (d, 1H, J=8.1); Anal. (C₃₀H₄₂FN₃O₆S.0.25H₂O) C, H,N.

Example 15

[0329] Preparation of Compound 15:Ethyl-3-(EtSCO-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-Gln)-E-Propenoate.

[0330] Preparation of Intermediatetrans-6-Methyl-2S-(4-trifluoromethyl-benzyl)-hept-4-enoic Acid(2R-Hydroxy-1R-methyl-2-phenyl-ethyl)-methyl Amide.

[0331] n-Butyllithium (25.5 mL of a 1.6 M solution in hexanes, 40.8mmol, 2.15 equiv) was added to a suspension of anhydrous lithiumchloride (5.64 g, 133 mmol, 7 equiv) and diisopropylamine (6.13 mL, 43.7mmol, 2.3 equiv) in THF (200 mL) at −78° C. The reaction mixture wasstirred for 30 min at −78° C., then maintained at 0° C. for 5 min, andsubsequently cooled again to −78° C. trans-6-Methyl-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (5.5 g, 19.0 mmol, 1equiv) in THF (40 mL) was added via cannula, and the resulting solutionwas stirred at −78° C. for 1.75 h, maintained at 0° C. for 20 min,stirred at 23° C. for 5 min, and then cooled again to 0° C. A solutionof 4-trifluoromethylbenzyl bromide (6.81 g, 28.5 mmol, 1.5 equiv) in THF(10 mL) was added, and the reaction mixture was stirred at 0° C. for 30min and then partitioned between half-saturated NH₄Cl (230 mL) and a 1:1mixture of EtOAc and hexanes (200 mL, 2×150 mL). The combined organiclayers were dried over Na₂SO₄ and concentrated. Purification of theresidue by flash column chromatography (gradient elution 20→40% EtOAc inhexanes) providedtrans-6-methyl-2S-(4-trifluoromethyl-benzyl)-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (7.64 g, 90%) as aviscous oil: R_(f)=0.47 (40% EtOAc in hexanes); IR (cm⁻¹) 3378, 1619; ¹HNGR (CDCl₃, mixture of rotamers) δ 0.85 (d, J=6.5), 0.89 (d, J=6.5),0.91 (d, J=6.5), 0.96 (d, J=6.8), 2.07-2.34 (m), 2.36-2.47 (m), 2.59(s), 2.76-2.86 (m), 2.88-3.01 (m), 3.07-3.22 (m), 3.96 (br), 3.99-4.09(m), 4.37-4.52 (m), 5.19-5.57 (m), 7.19-7.40 (m), 7.47-7.57 (m).

[0332] Preparation of Intermediate5S-(1R-Bromo-2-methyl-propyl)-3R-(4-trifluoromethyl-benzyl)-dihydrofuran-2-one.

[0333] N-Bromosuccinimide (3.17 g, 17.8 mmol, 1.05 equiv) was added insmall portions over 10 min to a solution oftrans-6-methyl-2S-(4-trifluoromethyl-benzyl)-hept-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (7.58 g, 16.9 mmol, 1equiv) and glacial acetic acid (4.85 mL, 84.7 mmol, 5 equiv) in a 4:1mixture of THF and H₂O (180 mL) at 0° C. The resulting yellow solutionwas stirred for 15 min at 0° C., then warmed to 23° C., and subsequentlyrefluxed for 45 min. After cooling to 23° C., the reaction mixture waspartitioned between half-saturated NaHCO₃ (200 mL) and a 1:1 mixture ofEtOAc and hexanes (2×200 mL, 100 mL). The combined organic layers weredried over Na₂SO₄ and concentrated. Flash chromatographic purificationof the residue (gradient elution 5→9→12% EtOAc in hexanes) gave5S-(1R-bromo-2-methyl-propyl)-3R-(4-trifluoromethyl-benzyl)-dihydrofuran-2-one(5.28 g, 82%) as a white solid (containing approximately 5-10%unidentified impurities by ¹H NGR): mp=83-85° C.; R_(f)=0.60 (25% EtOAcin hexanes); IR (cm⁻¹) 1778; ¹H NGR (CDCl₃, major isomer) δ 0.95 (d, 3H,J=6.5), 1.01 (d, 3H, J=6.8), 2.05-2.25 (m, 2H), 2.28-2.38 (m, 1H), 2.89(dd, 1H, J=13.5, 8.9), 2.98-3.09 (m, 1H), 3.22 (dd, 1H, J=13.5, 4.5),3.91 (dd, 1H, J=8.9, 3.6), 4.40-4.49 (m, 1H), 7.34 (d, 2H, J=8.1), 7.59(d, 2H, J=8.1); Anal. (C₁₆H₁₈BrF₃O₂) C, H.

[0334] Preparation of Intermediate5S-(1S-Azido-2-methyl-propyl)-3R-(4-trifluoromethyl-benzyl)-dihydrofuran-2-one.

[0335] A suspension of sodium azide (2.22 g, 34.1 mmol, 2.5 equiv) and5S-(1R-bromo-2-methyl-propyl)-3R-(4-trifluoromethyl-benzyl)-dihydrofuran-2-one(5.18 g, 13.7 mmol, 1 equiv) in N,N-dimethylformamide (50 mL) was heatedat 50° C. for 66 hours. The reaction mixture was cooled to 23° C. andpartitioned between half-saturated NaCl (200 mL) and a 1:1 mixture ofEtOAc and hexanes (3×200 mL). The combined organic layers were driedover Na₂SO₄, concentrated, and the residue purified by flash columnchromatography (gradient elution 9→12→20% EtOAc in hexanes) to give5S-(1S-azido-2-methyl-propyl)-3R-(4-trifluoromethyl-benzyl)-dihydrofuran-2-one(2.77 g, 59%) as a viscous oil (containing approximately 5-10%unidentified impurities by ¹H NGR): R_(f)=0.42 (25% EtOAc in hexanes);IR (cm⁻¹) 2097, 1772; ¹H NGR (CDCl₃, major isomer) δ 1.01 (d, 3H,J=6.5), 1.02 (d, 3H, J=6.8), 1.96-2.23 (m, 3H), 2.84-2.98 (m, 2H),3.09-3.20 (m, 1H), 3.25 (dd, 1H, J=13.7, 4.7), 4.44-4.52 (m, 1H), 7.34(d, 2H, J=8.1), 7.59 (d, 2H, J=8.1); Anal. (C₁₆H₁₈F₃N₃O₂) C, H, N.

[0336] Preparation of Intermediate{2-Methyl-1S-[4R-(4-trifluoromethyl-benzyl)-5-oxo-tetrahydrofuran-2S-yl]-propyl}-carbamicAcid tert-Butyl Ester.

[0337] A suspension of5S-(1S-azido-2-methyl-propyl)-3R-(4-trifluoromethyl-benzyl)-dihydrofuran-2-one(2.71 g, 7.94 mmol, 1 equiv), di-tert-butyl dicarbonate (2.43 g, 11.1mmol, 1.4 equiv) and Pd/C (10%, 0.225 g) in CH₃OH (110 mL) was stirredunder a hydrogen atmosphere (balloon) for 5 hours. The reaction mixturewas vacuum filtered through Whatman #5 paper and concentrated.Purification of the residue by flash column chromatography (gradientelution 12→20% EtOAc in hexanes) provided{2-methyl-1S-[4R-(4-trifluoromethyl-benzyl)-5-oxo-tetrahydrofuran-2S-yl]-propyl}-carbamicacid tert-butyl ester (1.87 g, 57%) as a white foam: R_(f)=0.84 (5% MeOHin CH₂Cl₂); IR (cm⁻¹) 3331, 1766, 1708, 1690; ¹H NGR (CDCl₃) δ 0.94 (d,3H, J=6.8), 0.96 (d, 3H, J=6.5), 1.40 (s, 9H), 1.71-1.86 (m, 1H),1.95-2.06 (m, 1H), 2.20-2.31 (m, 1H), 2.86 (dd, 1H, J=13.5, 8.9),2.93-3.04 (m, 1H), 3.20 (dd, 1H, J=13.5, 4.2), 3.33-3.42 (m, 1H),4.41-4.56 (m, 2H), 7.31 (d, 2H, J=8.1), 7.56 (d, 2H, J=8.1); Anal.(C₂₁H₂₈F₃NO₄) C, H, N.

[0338] Preparation of IntermediateEthyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-(Tr-Gln)]-E-Propenoate.

[0339] Lithium hydroxide (8.8 mL of a 1 M aqueous solution, 8.8 mmol, 5equiv) was added to a solution of{2-methyl-1S-[4R-(4-trifluoromethyl-benzyl)-5-oxo-tetrahydrofuran-2S-yl]-propyl}-carbamicacid tert-butyl ester (0.731 g, 1.76 mmol, 1 equiv) in DME (25 mL) at23° C. The resulting suspension was stirred at 23° C. for 30 min andthen partitioned between 10% KHSO₄ (35 mL) and CH₂Cl₂ (2×100 mL, 70 mL).The combined organic layers were dried over Na₂SO₄, concentrated, andthe residue dissolved in CH₂Cl₂/CH₃CN 10:1 (33 mL). Powdered 4 Åmolecular sieves (0.65 g), 4-methylmorpholine N-oxide (0.412 g, 3.52mmol, 2 equiv), and tetrapropylammonium perruthenate (0.062 g, 0.18mmol, 0.10 equiv) were added sequentially. The resulting dark reactionmixture was stirred for 2 hours at 23° C., and then vacuum filteredthrough Whatman #5 paper. The filtrate was then transferred to a flaskcontaining crude ethyl-3-[H₂N-L-(Tr-Gln)]-E-propenoate•HCl (2.12 mmol,1.2 equiv, prepared as described in Example 1 for the preparation ofethyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-propenoate),4-methylmorpholine (0.774 mL, 7.04 mmol, 4 equiv),1-hydroxybenzotriazole hydrate (0.357 g, 2.64 mmol, 1.5 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.506 g,2.64 mmol, 1.5 equiv) were added sequentially, and the reaction mixturewas stirred for 17 hours at 23° C. This mixture then was partitionedbetween brine (100 mL) and EtOAc (3×100 mL). The combined organic layerswere dried over Na₂SO₄ and concentrated. Purification of the residue byflash column chromatography (40% EtOAc in hexanes) providedethyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-(Tr-Gln)]-E-propenoate (0.839g, 56%) as a tan foam: R_(f)=0.27 (40% EtOAc in hexanes); IR (cm⁻¹)3308, 1711, 1666; ¹H NGR (CDCl₃) δ 0.67 (d, 3H, J=6.5), 0.92 (d, 3H,J=6.5), 1.27 (t, 3H, J=7.2), 1.39 (s, 9H), 1.58-1.71 (m, 1H), 1.90-2.06(m, 2H), 2.35-2.50 (m, 3H), 2.65-2.84 (m, 2H), 2.89-3.07 (m, 2H),4.03-4.22 (m, 3H), 4.38-4.45 (m, 1H), 4.86 (d, 1H, J=8.1), 5.51 (d, 1H,J=15.6), 6.32 (d, 1H, J=8.1), 6.62 (dd, 1H, J=15.6, 5.0), 7.10 (s, 1H),7.17-7.31 (m, 17H), 7.53 (d, 2H, J=7.8); Anal. (C₄₉H₅₆F₃N₃O₇.0.5 H₂O) C,H, N.

[0340] Preparation of IntermediateEthyl-3-[EtSCO-L-ValΨ[COCH₂]-L-p-CF₃)Phe-L-(Tr-Gln)]-E-Propenoate.

[0341] HCl (3 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution ofethyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-(Tr-Gln)]-E-propenoate (0.271g, 0.317 mmol) in 1,4-dioxane (3 mL). The reaction solution was stirredat 23° C. for 2 hours and then concentrated. The residue was dissolvedin dry CH₂Cl₂ (5 mL). N,N-diisopropylethylamine (0.165 mL, 0.947 mmol, 3equiv) and ethyl chlorothiolformate (0.056 mL, 0.54 mmol, 1.7 equiv)were added sequentially. The reaction solution was stirred 2 hours at23° C. and then partitioned between brine (30 mL) and CH₂Cl₂ (3×30 mL).The combined organic phases were dried over Na₂SO₄,concentrated, and theresidue was chromatographed on silica gel (40% EtOAc in hexanes) toaffordethyl-3-[EtSCO-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-(Tr-Gln)]-E-propenoate(0.142 g, 53%) as a white foam: R_(f)=0.25 (40% EtOAc in hexanes); IR(cm⁻¹) 3304, 1713, 1653; ¹H NGR (CDCl₃) δ 0.70 (d, 3H, J=6.8), 0.94 (d,3H, J=6.5), 1.24 (t, 3H, J=7.5), 1.27 (t, 3H, J=7.2), 1.59-1.72 (m, 1H),1.91-2.04 (m, 2H), 2.33-2.51 (m, 3H), 2.65-3.08 (m, 6H), 4.08-4.22 (m,2H), 4.34-4.48 (m, 2H), 5.52 (dd, 1H, J=15.7, 1.7), 5.66 (d, 1H, J=8.1),6.39 (d, 1H, J=7.8), 6.62 (dd, 1H, J=15.7, 5.1), 7.04 (s, 1H), 7.17-7.30(m, 17H), 7.53 (d, 2H, J=7.8).

[0342] Preparation of ProductEthyl-3-(EtSCO-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-Gln)-E-Propenoate.

[0343]Ethyl-3-[EtSCO-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-(Tr-Gln)]-E-propenoate(0.112 g, 0.133 mmol, 1 equiv) was dissolved in dry CH₂Cl₂ (6 mL).Triisopropylsilane (0.082 mL, 0.40 mmol, 3 equiv) and trifluoroaceticacid (3 mL) were added sequentially producing a bright yellow solution.This solution was stirred for 40 minutes and then concentrated. Theresidue was stirred in Et₂O (6 mL), and the solid was collected byfiltration, washed with Et₂O (2×3 mL), and then dried under vacuum togive ethyl-3-(EtSCO-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-Gln)-E-propenoate(0.068 g, 85%) as a white solid: mp=216° C. (dec); R_(f)=0.43 (10% CH₃OHin CH₂Cl₂); IR (cm⁻¹) 3425, 3308, 1717, 1655; ¹H NGR (DMSO-d₆) δ 0.76(d, 3H, J=6.5), 0.82 (d, 3H, J=6.5), 1.12-1.23 (m, 6H), 1.53-1.75 (m,2H), 1.99-2.14 (m, 3H), 2.50-2.60 (m, 1H), 2.64-2.87 (m, 5H), 2.93-3.04(m, 1H), 3.98-4.21 (m, 3H), 4.26-4.36 (m, 1H), 5.50 (d, 1H, J=15.5),6.62 (dd, 1H, J=15.5, 5.4), 6.74 (s, 1H), 7.15 (s, 1H), 7.37 (d, 2H,J=8.1), 7.59 (d, 2H, J=8.1), 8.06 (d, 1H, J=8.1), 8.35 (d, 1H, J=8.1);Anal. (C₂₈H₃₈F₃N₃O₆S) C, H, N.

Example 16

[0344] Preparation of Compound 16:Ethyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CF3)Phe-L-Gln)-E-Propenoate.

[0345] Preparation of IntermediateEthyl-3-[CyPentylSCO-L-ValΨ[COCH₂]-L-p-CF₃)Phe-L-(Tr-Gln)]-E-Propenoate.

[0346] HCl (3 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution ofethyl-3-[BOC-L-ValΨ[COCH₂]-L-p-CF₃)Phe-L-(Tr-Gln)]-E-propenoate (0.268g, 0.313 mmol) in 1,4-dioxane (3 mL). The reaction solution was stirredat 23° C. for 2 hours and then concentrated. The residue was dissolvedin dry CH₂Cl₂ (5 mL). N,N-diisopropylethylamine (0.164 mL, 0.941 mmol, 3equiv) and cyclopentyl chlorothiolformate (0.088 mL, 1.7 equiv) wereadded sequentially. The reaction solution was stirred 3 hours at 23° C.and then was partitioned between brine (30 mL) and CH₂Cl₂ (3×30 mL). Thecombined organic phases were dried over Na₂SO₄, concentrated, and theresidue was chromatographed on silica gel (40% EtOAc in hexanes) toaffordethyl-3-[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-(Tr-Gln)]-E-propenoate(0.169 g, 61%) as a white foam: R_(f)=0.25 (40% EtOAc in hexanes); IR(cm⁻¹) 3324, 1718, 1657; ¹H NGR (CDCl₃) δ 0.70 (d, 3H, J=6.8), 0.93 (d,3H, J=6.5), 1.27 (t, 3H, J=7.2), 1.46-1.73 (m, 7H), 1.91-2.12 (m, 4H),2.30-2.52 (m, 3H), 2.65-2.83 (m, 2H), 2.90-3.06 (m, 2H), 3.56-3.66 (m,1H), 4.08-4.22 (m, 2H), 4.31-4.49 (m, 2H), 5.51 (dd, 1H, J=15.8, 1.6),5.65 (d, 1H, J=7.8), 6.40 (d, 1H, J=7.8), 6.62 (dd, 1H, J=15.8, 5.1),7.08 (s, 1H), 7.17-7.30 (m, 17H), 7.53 (d, 2H, J=8.1); Anal.(C₅₀H₅₆F₃N₃O₆S.O.5 H₂O) C, H, N.

[0347] Preparation of ProductEthyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-Gln)-E-Propenoate.

[0348]Ethyl-3-[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-(Tr-Gln)]-E-propenoate(0.148 g, 0.167 mmol, 1 equiv) was dissolved in dry CH₂Cl₂ (6 mL).Triisopropylsilane (0.103 mL, 0.503 mmol, 3 equiv) and trifluoroaceticacid (3 mL) were added sequentially producing a bright yellow solution.This solution was stirred for 40 minutes and then concentrated. Theresidue was stirred in Et₂O (6 mL), and the solid was collected byfiltration, washed with Et₂O (2×3 mL), and then dried under vacuum togive ethyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CF₃)Phe-L-Gln)-E-propenoate(0.089 g, 83%) as a white solid: mp=225° C. (dec); R_(f)=0.44 (10% CH₃OHin CH₂Cl₂); IR (cm⁻¹) 3425, 3303, 1717, 1654; ¹H NGR (DMSO-d₆) δ 0.76(d, 3H, J=6.5), 0.82 (d, 3H, J=6.8), 1.21 (t, 3H, J=7.2), 1.37-1.74 (m,8H), 1.92-2.13 (m, SH), 2.50-2.59 (m, 1H), 2.64-2.87 (m, 3H), 2.93-3.04(m, 1H), 3.46-3.57 (m, 1H), 4.02-4.18 (m, 3H), 4.25-4.36 (m, 1H), 5.51(dd, 1H, J=15.8, 1.2), 6.62 (dd, 1H, J=15.8, 5.4), 6.74 (s, 1H), 7.16(s, 1H), 7.37 (d, 2H, J=8.1), 7.59 (d, 2H, J=8.1), 8.06 (d, 1H, J=8.4),8.28 (d, 1H, J=7.8); Anal. (C₃₁H₄₂F₃N₃O₆S) C, H, N.

Example 17

[0349] Preparation of Compound 17:[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln]-E-1-Acetyl-3-methylene-pyrrolidin-2-one.

[0350] Preparation of Intermediate[1-(1-Acetyl-2-oxo-pyrrolidin-3-ylidenemethyl)-3-(S)-(trityl-carbamoyl)-propyl]-carbamicAcid tert-Butyl Ester.

[0351] Triphenylphosphine (0.646 g, 2.46 mmol, 1.40 equiv) was added toa solution of 1-acetyl-3-bromo-pyrrolidin-2-one (prepared as describedin: Ikuta, H., Shirota, H., Kobayashi, S., Yamagishi, Y., Yamada, K.,Yamatsu, I., Katayama, K., J. Med. Chem. 1987, 30, 1995, which documentis entirely incorporated herein by reference) (0.378 g, 1.76 mmol, 1.0equiv) in THF (10 mL). The reaction mixture was refluxed for 5 h, andthen the solvent was evaporated to give the crude salt. This materialwas dissolved in EtOH (10 mL), and Et₃N (0.613 mL, 4.4 mmol, 2.5 equiv)and [BOC-L-(Tr-Gln)]-H (0.832 g, 1.76 mmol, 1 equiv) were addedsequentially. The reaction mixture was stirred at 60° C. for 24 h, thenthe volatiles were evaporated, and the residue was partitioned betweenCH₂Cl₂ (50 mL) and brine (50 mL). The organic layer was dried overNa₂SO₄ and concentrated. The residue was purified by flashchromatography on silica gel (35% EtOAc in hexanes) to afford[1-(1-acetyl-2-oxo-pyrrolidin-3-ylidenemethyl)-3-(S)-(trityl-carbamoyl)-propyl]-carbamicacid tert-butyl ester as a pale yellow foam (0.381 g, 38%): R_(f)=0.30(33% EtOAc in hexanes); IR (cm⁻¹) 1687, 1510, 1366, 1274; ¹H NGR (CDCl₃)δ1.42 (s, 9H), 1.80-1.89 (m, 2H), 2.35-2.44 (m, 2H), 2.55 (s, 3H),2.57-2.64 (m, 1H), 2.77-2.82 (m, 1H), 3.72-3.77 (m, 2H), 4.25 (m, 1H),4.80 (d, 1H, J=8.1), 6.40-6.44 (m, 1H), 6.82 (s, br, 1H), 7.20-7.33 (m,15H); Anal. (C₃₅H₃₉N₃O₅.1.0 H₂O) C, H, N.

[0352] Preparation of Intermediate[BOC-L-ValΨ[COCH₂]-L-p-CH₃)Phe-L-(Tr-Gln)]-E-1-Acetyl-3-methylene-pyrrolidin-2-one.

[0353] HCl (5 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of[1-(-acetyl-2-oxo-pyrrolidin-3-ylidenemethyl)-3-S-(trityl-carbamoyl)-propyl]-carbamicacid tert-butyl ester (0.442 g, 0.760 mmol) in 1,4-dioxane (5 mL). Thereaction solution was stirred at 23° C. for 2 hours. It then wasconcentrated and set aside.

[0354] Lithium hydroxide (3.46 mL of a 1 M aqueous solution, 3.46 mmol,5 equiv) was added to a solution of{2-methyl-1S-[4R-(4-methyl-benzyl)-5-oxo-tetrahydrofuran-2S-yl]-propyl}-carbamicacid tert-butyl ester (0.250 g, 0.692 mmol, 1 equiv) in DME (10 mL) at23° C. The resulting suspension was stirred at 23° C. for 30 min andthen partitioned between 10% KHSO₄ (20 mL) and CH₂Cl₂ (3×70 mL). Thecombined organic layers were dried over Na₂SO₄, concentrated, and theresidue dissolved in CH₂Cl₂ (12 mL). Powdered 4 Å molecular sieves (0.25g), 4-methylmorpholine N-oxide (0.162 g, 1.38 mmol, 2 equiv), andtetrapropylammonium perruthenate (0.024 g, 0.068 mmol, 0.10 equiv) wereadded sequentially. The resulting dark reaction mixture was stirred for1.5 hours at 23° C. and then vacuum filtered through Whatman #5 paper.The filtrate was added directly to the crude amine salt prepared above.4-Methylmorpholine (0.304 mL, 2.77 mmol, 4 equiv),1-hydroxybenzotriazole hydrate (0.140 g, 1.04 mmol, 1.5 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.199 g,1.04 mmol, 1.5 equiv) were added sequentially, and the reaction mixturewas stirred for 19 hours at 23° C. and then loaded directly onto a flashcolumn for chromatographic purification (50% EtOAc in hexanes) providing[BOC-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-1-acetyl-3-methylene-pyrrolidin-2-one(0.233 g, 40%) as a white foam: R_(f)=0.25 (50% EtOAc in hexanes); IR(cm⁻¹) 3336, 1687; ¹H NGR (CDCl₃) δ 0.60 (d, 3H, J=6.8), 0.91 (d, 3H,J=6.8), 1.42 (s, 9H), 1.55-1.68 (m, 2H), 1.82-1.96 (m, 2H), 2.30 (s,3H), 2.34-2.83 (m, 7H), 2.55 (s, 3H), 2.87-2.98 (m, 1H), 3.64-3.72 (m,2H), 4.03-4.08 (m, 1H), 4.26-4.38 (m, 1H), 4.78 (d, 1H, J=8.1), 5.79 (d,1H, J=8.1), 6.10-6.16 (m, 1H), 6.96 (d, 2H, J=7.9), 7.06 (d, 2H, J=7.9),7.17-7.33 (m, 16H).

[0355] Preparation of Intermediate[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-1-Acetyl-3-methylene-pyrrolidin-2-one.

[0356] HCl (3 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of[BOC-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-1-acetyl-3-methylene-pyrrolidin-2-one(0.217 g, 0.258 mmol) in 1,4-dioxane (3 mL). The reaction solution wasstirred at 23° C. for 2 hours and then concentrated. The residue wasdissolved in dry CH₂Cl₂ (5 mL). N,N-diisopropylethylamine (0.135 mL,0.775 mmol, 3 equiv) and cyclopentyl chlorothiolformate (0.072 mL, 1.7equiv) were added sequentially. The reaction solution was stirred 3hours at 23° C. and then partitioned between brine (30 mL) and CH₂Cl₂(3×30 mL). The combined organic phases were dried over Na₂SO₄,concentrated, and the residue was chromatographed on silica gel (50%EtOAc in hexanes) to afford[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-1-acetyl-3-methylene-pyrrolidin-2-one(0.114 g, 51%) as a colorless glass: R_(f)=0.26 (50% EtOAc in hexanes);IR (cm⁻¹) 3328, 1719, 1670; ¹H NGR (CDCl₃) δ 0.64 (d, 3H, J=6.8), 0.92(d, 3H, J=6.5), 1.47-1.78 (m, 8H), 1.80-1.95 (m, 2H), 1.99-2.17 (m, 2H),2.29-2.84 (m, 7H), 2.30 (s, 3H), 2.55 (s, 3H), 2.94 (dd, 1H, J=17.4,10.0), 3.59-3.73 (m, 3H), 4.25-4.38 (m, 2H), 5.57 (d, 1H, J=8.1), 5.89(d, 1H, J=8.1), 6.11-6.17 (m, 1H), 6.96 (d, 2H, J=7.9), 7.06 (d, 2H,J=7.9), 7.18-7.33 (m, 16H); Anal. (C₅₂H₆₀N₄O₆S) C, H, N.

[0357] Preparation of Product[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln]-E-1-Acetyl-3-methylene-pyrrolidin-2-one.

[0358][CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-1-acetyl-3-methylene-pyrrolidin-2-one(0.093 g, 0.107 mmol, 1 equiv) was dissolved in dry CH₂Cl₂ (6 mL).Triisopropylsilane (0.066 mL, 0.32 mmol, 3 equiv) and trifluoroaceticacid (3 mL) were added sequentially producing a bright yellow solution.This solution was stirred for 40 minutes and then concentrated. Theresidue was stirred in Et₂O (6 mL), and the solid was collected byfiltration, washed with Et₂O (2×3 mL), and then dried under vacuum togive[CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln]-E-1-acetyl-3-methylene-pyrrolidin-2-one(0.049 g, 73%) as a white solid: mp=200° C. (dec); R_(f)=0.47 (10% CH₃OHin CH₂Cl₂); IR (cm⁻¹) 3425, 3337, 1719, 1670; ¹H NGR (DMSO-d₆) δ 0.74(d, 3H, J=6.5), 0.81 (d, 3H, J=6.5), 1.36-1.79 (m, 8H), 1.89-2.13 (m,5H), 2.20 (s, 3H), 2.39-2.90 (m, 7H), 2.43 (s, 3H), 3.46-3.64 (m, 3H),4.10-4.16 (m, 1H), 4.24-4.36 (m, 1H), 6.12-6.18 (m, 1H), 6.74 (s, 1H),6.99 (s, 4H), 7.15 (s, 1H), 8.05 (d, 1H, J=7.8), 8.25 (d, 1H, J=7.8);Anal. (C₃₃H₄₆N₄O₆S.0.5 H₂O) C, H, N.

Example 18

[0359] Preparation of Compound 18:Ethyl-3-(CyPentylSCO-L-PheΨ[COCH₂]-L-(p-F)Phe-L-Gln)-E-Propenoate.

[0360] Preparation of Intermediate 3-(4-Fluoro-phenyl)-propionic Acidcis-1S-Amino-2R-indanol-acetonide Amide.

[0361] Oxalyl chloride (6.14 mL, 70.4 mmol, 1.05 equiv) was added to asolution of 3-(4-fluoro-phenyl)-propionic acid (11.3 g, 67.2 mmol, 1equiv) and N,N-dimethylformamide (0.03 mL, 0.39 mmol, 0.006 equiv) inbenzene (150 mL) at 23° C. The reaction mixture was stirred at 23° C.for 1.5 h and then concentrated. The resulting oil was dissolved in THF(30 mL) and was added to a 0° C. solution of(1S,2R)-cis-1-amino-2-indanol (10.0 g, 67.0 mmol, 1.0 equiv) and Et₃N(10.3 mL, 73.9 mmol, 1.1 equiv) in THF (250 mL). After stirring for 20min at 0° C., the reaction mixture was partitioned betweenhalf-saturated NH₄Cl (150 mL) and EtOAc (2×150 mL). The organic layerswere dried over Na₂SO₄ and concentrated to afford a white solid. Thismaterial was dissolved in a mixture of CH₂Cl₂ (400 mL) and2-methoxypropene (30 mL), and the resulting solution was treated withmethanesulfonic acid (0.20 mL). After stirring 15 min at 23° C., thereaction mixture was partitioned between half-saturated NaHCO₃ (150 mL)and CH₂Cl₂ (2×150 mL). The combined organic layers were dried over MgSO₄and gravity filtered. The filtrate was concentrated, and the residue waspurified by flash column chromatography (gradient elution, 10→20% EtOAcin hexanes) to provide 3-(4-fluoro-phenyl)-propionic acidcis-1S-amino-2R-indanol-acetonide amide (18.2 g, 83%) as a pale yellowoil: R_(f)=0.52 (50% EtOAc in hexanes); IR (cm⁻¹) 2934, 1645; ¹H NGR(CDCl₃) δ 1.34 (s, 3H), 1.60 (s, 3H), 2.91-2.95 (m, 2H), 3.04-3.13 (m,4H), 4.68-4.71 (m, 1H), 5.06 (d, 1H, J=4.7), 6.94-7.00 (m, 2H),7.02-7.30 (m, 6H); Anal. (C₂₁H₂₂FNO₂) C, H, N.

[0362] Preparation of Intermediate{1S-[4R-(4-Fluoro-benzyl)-5-oxo-tetrahydro-furan-2S-yl]-2-phenyl-ethyl}-carbamicAcid tert-Butyl Ester.

[0363] n-Butyllithium (13.5 mL of a 1.6 M solution in hexanes, 21.6mmol, 2.0 equiv) was added to a solution of(1R-oxiranyl-2S-phenyl-ethyl)-carbamic acid tert-butyl ester (preparedaccording to Luly, J. R., Dellaria, J. F., Plattner, J. J., Soderquist,J, L., Yi, N., J. Org. Chem. 1987, 52, 1487) (2.85 g, 10.8 mmol, 1equiv) and 3-(4-fluoro-phenyl)-propionic acidcis-1S-amino-2R-indanol-acetonide amide (3.67 g, 10.8 mmol, 1.0 equiv)in THF (150 mL) at −78° C. The reaction mixture was stirred for 5 min at−78° C., maintained at 0° C. for 1 h, and then partitioned between 0.5 MHCl (150 mL) and a 1:1 mixture of EtOAc and hexanes (2×150 mL). Theorganic layers were dried over Na₂SO₄ and concentrated. Flashchromatographic purification of the residue (gradient elution, 25→40%EtOAc in hexanes) gave the coupling product (3.19 g, 49%) as a yellowoil contaminated with several minor impurities. This material wasdissolved in a 5:1 mixture of toluene and CH₂Cl₂ (180 mL) and treatedwith p-toluenesulfonic acid monohydrate (1.01 g, 5.31 mmol, 1.0 equiv)at 23° C. After stirring for 13 h at 23° C., the reaction mixture wasfiltered through a medium frit, and the filtrate was partitioned betweenhalf-saturated NaHCO₃ (150 mL) and a 1:1 mixture of EtOAc and hexanes(2×150 mL). The organic layers were dried over Na₂SO₄ and concentrated.The residue was purified by flash column chromatography (20% EtOAc inhexanes) to provide{1S-[4R-(4-fluoro-benzyl)-5-oxo-tetrahydro-furan-2S-yl]-2-phenyl-ethyl}-carbamicacid tert-butyl ester (1.28 g, 59%) as a white foam: R_(f)=0.46 (30%EtOAc in hexanes); IR (cm⁻¹) 3332, 2976, 1767, 1702; ¹H NGR (CDCl₃) δ1.36 (s, 9H), 1.88-1.97 (m, 1H), 2.19-2.29 (m, 1H), 2.75-2.99 (m, 4H),3.05 (dd, 1H, J=13.5, 4.5), 3.93 (q, 2H, J=8.5), 4.13-4.18 (m, 1H), 4.54(d, 1H, J=9.7), 6.91-6.98 (m, 2H), 7.08-7.32 (m, 7H); Anal. (C₂₄H₂₈FNO₄)C, H, N

[0364] Preparation of IntermediateEthyl-3-[BOC-L-PheΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-Propenoate.

[0365] Lithium hydroxide (7.6 mL of a 1 M aqueous solution, 7.6 mmol, 5equiv) was added to a solution of{1S-[4R-(4-fluoro-benzyl)-5-oxo-tetrahydro-furan-2S-yl]-2-phenyl-ethyl}-carbamicacid tert-butyl ester (0.630 g, 1.52 mmol, 1 equiv) in DME (8 mL) at 23°C. The resulting suspension was stirred at 23° C. for 20 min and thenpartitioned between 0.5 M HCl (100 mL) and EtOAc (2×100 mL). Thecombined organic layers were dried over Na₂SO₄, concentrated, and theresidue dissolved in a 1:1 mixture of CH₂Cl₂ and CH₃CN (100 mL).4-Methylmorpholine N-oxide (0.357 g, 3.05 mmol, 2 equiv), powdered 4 Åmolecular sieves (0.70 g), and tetrapropylammonium perruthenate (0.054g, 0.153 mmol, 0.1 equiv) were added sequentially. The resulting darkreaction mixture was stirred for 3 h at 23° C. and then filtered throughcelite. The filtrate was concentrated under reduced pressure to providea brown oil which was dissolved in CH₂Cl₂ (40 mL). Crudeethyl-3-[H₂N-L-(Tr-Gln)]-E-propenoate•HCl (see preparation ofethyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-propenoate, 1.27 mmol,1.2 equiv), 1-hydroxybenzotriazole hydrate (0.268 g, 1.98 mmol, 1.3equiv), 4-methylmorpholine (0.670 mL, 6.09 mmol, 4 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.380 g,1.98 mmol, 1.3 equiv) were added sequentially, and the reaction mixturewas stirred for 22 h at 23° C. and then partitioned between water (150mL) and a 1:1 mixture of EtOAc and hexanes (2×150 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated. Purification ofthe residue by flash column chromatography (40% EtOAc in hexanes)provided ethyl-3-[BOC-L-PheΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.558 g, 43%) as a white solid: mp=89-100° C.; R_(f)=0.44 (50% EtOAc inhexanes); IR (cm⁻¹) 3316, 2972, 1708, 1665; ¹H NGR (CDCl₃) δ 1.29 (t,3H, J=7.2), 1.35 (s, 9H), 1.95-2.05 (m, 1H), 2.34-2.39 (m, 2H), 2.46 (d,1H, J=16.8), 2.57-2.99 (m, 7H), 4.17 (q, 2H, J=7.2), 4.27-4.33 (m, 1H),4.48 (s, br, 1H), 4.58 (d, 1H, J=6.9), 5.42 (d, 1H, J=15.3), 6.08 (d,1H, J=8.4), 6.62 (dd, 1H, J=15.3, 4.8), 6.93-7.19 (m, 6H), 7.21-7.29 (m,19H); Anal. (C₅₂H₅₆FN₃O₇) C, H, N.

[0366] Preparation of IntermediateEthyl-3-[CyPentylSCO-L-PheΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-Propenoate.

[0367] HCl (8 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution ofethyl-3-[BOC-L-PheΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate (0.302 g,0.354 mmol, 1 equiv) in 1,4-dioxane (10 mL). The reaction mixture wasstirred at 23° C. for 1.5 h and then concentrated. The resulting oil wasdissolved in CH₂Cl₂ (15 mL), cooled to ⁰° C., and 4-methylmorpholine(0.117 mL, 1.06 mmol, 3.0 equiv) and cyclopentyl chlorothiolformate(0.087 mL, 0.528 mmol, 1.5 equiv) were added sequentially. The reactionmixture was stirred for 30 min at 0° C. and then partitioned betweenwater (100 mL) and a 1:1 mixture of EtOAc and hexanes (2×100 mL). Thecombined organic layers were dried over Na₂SO₄, concentrated, and theresidue was chromatographed on silica gel (gradient elution, 30→40%EtOAc in hexanes) to affordethyl-3-[CyPentylSCO-L-PheΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.163 g, 52%) as a white solid: mp=75-85° C.; R_(f)=0.48 (50% EtOAc inhexanes); IR (cm⁻¹) 3314, 1710, 1655; ¹H NGR (CDCl₃) δ 1.29 (t, 3H,J=7.2), 1.48-1.67 (m, 6H), 1.97-2.03 (m, 2H), 2.29-2.42 (m, 2H),2.54-2.97 (m, 8H), 3.54-3.63 (m, 1H), 4.18 (q, 2H, J=7.2), 4.51-4.58 (m,2H), 5.44 (dd, 1H, J=15.6, 1.7), 5.59 (d, 1H, J=6.9), 5.90 (d, 1H,J=7.2), 6.16 (d, 1H, J=8.4), 6.64 (dd, 1H, J=15.6, 5.0), 6.91-7.08 (m,5H), 7.14-7.29 (m, 20H); Anal. (C₅₃H₅₆FN₃O₆S) C, H, N.

[0368] Preparation of ProductEthyl-3-(CyPentylSCO-L-PheΨ[COCH₂]-L-(p-F)Phe-L-Gin)-E-Propenoate.

[0369] Triisopropylsilane (0.10 mL) and trifluoroacetic acid (6 mL) wereadded sequentially to a solution ofethyl-3-[CyPentylSCO-L-PheΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.160 g, 0.181 mmol) in CH₂Cl₂ (10 mL) producing a bright yellowsolution. The reaction mixture was stirred for 30 min at 23° C., thencarbon tetrachloride (6 mL) was added, and the mixture was concentratedunder reduced pressure. The residue was purified by flash columnchromatography (5% CH₃OH in CH₂Cl₂) to affordethyl-3-(CyPentylSCO-L-PheΨ[COCH₂]-L-(p-F)Phe-L-Gln)-E-propenoate (0.082g, 71%) as a white solid: mp=210-212° C.; R_(f)=0.10 (10% CH₃OH inCH₂Cl₂); IR (cm⁻¹) 3284, 1717, 1637; ¹H NGR (DMSO-d₆) δ 1.21 (t, 3H,J=6.8), 1.33-1.72 (m, 8H), 1.90-2.07 (m, 4H), 2.49-3.07 (m, 7H),3.43-3.47 (m, 1H), 4.09 (q, 2H, J=6.8), 4.33-4.35 (m, 2H), 5.37-5.46 (m,1H), 6.59-6.67 (m, 1H), 6.77 (s, br, 1H), 7.00-7.28 (m, 9H), 8.04 (d,1H, J=7.8), 8.46 (d, 1H, J=7.5), 8.53 (d, 1H, J=7.5); Anal.(C₃₂H₄₂FN₃O₆S) C, H, N.

Example 19

[0370] Preparation of Compound 19:Ethyl-3-(CyPentyl-L-LeuΨ[COCH₂]-L-p-F)Phe-L-Gln)-E-Propenoate.

[0371] Preparation of Intermediate BOC-L-Leucinol.

[0372] To a solution of BOC-Leu-OH (15.09 g, 65 mmol, 1 equiv) in THF(150 mL) at 0° C. was added BH₃. THF (163 mL of a 1.0 M solution in THF,163 mmol, 2.51 equiv). The reaction mixture was stirred at 23° C. for 3h and then cooled to 0° C. Brine (100 mL) was added carefully, and theaqueous layer was extracted with EtOAc (2×100 mL). The combined organiclayers were dried over Na₂SO₄ and concentrated. The residue was purifiedby flash column chromatography (gradient elution, 25→35% EtOAc inhexanes) to afford BOC-L-Leucinol as a colorless oil (12.59 g, 89%): IR(cm⁻¹): 3434, 1699, 1256, 739; ¹H NGR (CDCl₃) δ 0.93 (dd, 6H, J=6.6,1.5), 1.29-1.31 (m, 2H), 1.45 (s, 9H), 1.60-2.05 (m, 2H), 3.47-3.53 (m,1H), 3.65-3.72 (m, 2H), 4.56 (s, br, 1H); Anal. (C₁₁H₂₃NO₃) C, H, N.

[0373] Preparation of Intermediate BOC-L-Leucinal.

[0374] A solution of sulfur trioxide pyridine complex (7.8 g, 49.1 mmol,3.0 equiv) in a 1:1 mixture of DMSO and CH₂Cl₂ (100 mL) was added to asolution of BOC-L-Leucinol (3.56 g, 16.4 mmol. 1 equiv) and Et₃N (8 mL)in a 1:1 mixture of DMSO and CH₂Cl₂ (100 mL) at 0° C. The reactionmixture was stirred at 23° C. for 30 min, then poured into ice water(300 mL), and extracted with Et₂O (2×200 mL). The combined organiclayers were washed with 0.5 M HCl (150 mL), half saturated NaHCO₃ (150mL) and H₂O (2×150 mL), then dried over Na₂SO₄, and concentrated toafford crude BOC-L-Leucinal as pale yellow oil (3.48 g, 99%). Thismaterial was used without further purification. ¹H NGR (CDCl₃) δ0.92-0.94 (m, 6H), 1.44 (s, 9H), 1.59-1.64 (m, 1H), 1.67-1.85 (m, 2H),(4.21 (m, 1H), 5.12 (s, br, 1H), 9.57 (s, 1H).

[0375] Preparation of Intermediate [1S-Isobutyl-allyl]-carbamic Acidtert-Butyl Ester.

[0376] KN(TMS)₂ (67.9 mL of a 0.5 M solution in toluene, 33.9 mmol, 2.1equiv) was added dropwise to a solution of methyltriphenylphosphoniumbromide (12.1 g, 33.9 mmol, 2.1 equiv) in a 5:1 mixture of THF and DMSO(600 mL) at 0° C. The reaction mixture was stirred at 0° C. for 1 h,then cooled to −78° C., and a solution of BOC-L-Leucinal (3.48 g, 16.2mmol, 1 equiv) in THF (60 mL) was added. The reaction mixture wasstirred at −78° C. for 10 min and then warmed to 23° C. slowly. Themixture was partitioned between 0.5 M HCl (200 mL) and a 1:1 mixture ofEtOAc in hexanes (2×150 mL). The combined organic layers were washedwith H₂O (150 mL), dried over Na₂SO₄, and concentrated. The residue waspurified by flash column chromatography (10% EtOAc in hexanes) to afford(1S-isobutyl-allyl)-carbamic acid tert-butyl ester as a pale yellowsolid (2.44 g, 70%): R_(f)=0.78 (20% EtOAC in hexanes); IR (cm⁻¹): 3343,2958, 1594, 1520; ¹H NGR (CDCl₃) δ 0.92 (dd, 6H, J=6.6, 2.1), 1.30-1.38(m, 2H), 1.45 (s, 9H), 1.61-1.74 (m, 1H), 4.13 (m, 1H), 4.38 (s, br,1H), 5.08 (dd, 1H, J=10.2, 1.5), 5.15 (dd, 1H, J=17.4, 1.5), 5.73 (dd,1H, J=16.8, 10.2); Anal. (C₁₁H₂₃NO2-H₂O) C, H, N.

[0377] Preparation of Intermediate[3S-methyl-1R-oxiranyl-butyl]-carbamic Acid tert-Butyl Ester

[0378] 3-Chloroperoxybenzoic acid (˜60%, 8.40 g, ˜29.2 mmol, 1.8 equiv)was added to a solution of (1S-isobutyl-allyl)-carbamic acid tert-butylester (3.48 g, 16.2 mmol, 1 equiv) in CH₂Cl₂ (100 mL). The reactionmixture was stirred at 23° C. for 6 h, then poured into a 1:1 mixture ofEtOAc and hexanes (150 mL), and washed with 10% Na₂S₂O₅ (150 mL) andhalf-saturated NaHCO₃ (150 mL). The organic layer was dried over Na₂SO₄and concentrated. The residue was purified by flash columnchromatography (10% EtOAc in hexanes) to afford[3S-methyl-1R-oxiranyl-butyl]-carbamic acid tert-butyl ester as a paleyellow oil (3.15 g, 85%): R_(f)=0.55 (20% EtOAc in hexanes); IR (cm⁻¹)1702, 1501, 1366, 1168; ¹H NGR (CDCl₃) δ 0.96 (d, 6H, J=6.9),1.43 (s,9H), 1.64-1.81 (m, 1H), 2.60 (s, br, 1H), 2.72-2.76 (m, 2H), 2.84 (m,1H), 2.99 (s, br, 1H), 3.60 (s, br, 1H), 3.97-3.99 (m, 1H), 4.29 (m,1H).

[0379] Preparation of Intermediate{1S-[4R′-(4-Fluoro-benzyl)-5-oxo-tetrahydrofuran-2S′-yl]-3-methylbutyl}-carbamicAcid tert-Butyl Ester.

[0380] n-Butyllithium (8.79 mL of a 1.6 M solution in hexanes, 14.06mmol, 2.0 equiv) was added to a solution of[3S-methyl-1R-oxiranyl-butyl]-carbamic acid tert-butyl ester (1.61 g,7.03 mmol, 1.0 equiv) and 3-(4-fluoro-phenyl)-propionic acidcis-1S-amino-2R-indanol-acetonide amide (2.39 g, 7.03 mmol, 1 equiv) inTHF (100 mL) at −78° C. The resulting yellow solution was stirred at−78° C. for 5 min and then warmed to 0° C. for 1 h. The reaction wasquenched with 0.5 M HCl (100 mL) and extracted with a 1:1 mixture ofEtOAc in hexanes (2×100 mL). The combined organic layers were washedwith brine (150 mL), dried over Na₂SO₄, and concentrated. The residuewas purified by flash chromatography on silica gel (20% EtOAc inhexanes) to afford the coupling product as a white foam (2.28 g, 57%):R_(f)=0.44 (25% EtOAc in hexanes); IR (cm⁻¹) 3431, 1637, 1510, 1223; ¹HNGR (CDCl₃) δ (mixture of diastereomers) 0.92 (dd, J=6.6, 2.1),1.26-1.46 (m), 1.60 (s), 1.65 (s), 1.68-1.89 (m), 2.65-2.69 (m),2.75-2.81 (m), 2.91-2.96 (m), 3.06-3.13 (m), 3.24 (m), 3.34-3.48 (m),3.66 (m), 4.60 (d, J=7.5), 4.68-4.71 (m), 4.76-4.78 (m), 4.83 (s, br),5.06 (d, J=4.8), 6.22 (d, J=7.2), 6.91-7.31 (m).

[0381] To a solution of this material (2.21 g, 3.89 mmol, 1 equiv) in a5:1 mixture of toluene and CH₂Cl₂ (120 mL) was added p-toluenesulfonicacid monohydrate (0.739 g, 3.89 mmol, 1.0 equiv) at 23° C. The reactionmixture was stirred at 23° C. for 14 h and then quenched withhalf-saturated NaHCO₃ (100 mL). The resulting mixture was extracted witha 1:1 mixture of EtOAc in hexanes (2×100 mL), and the organic layerswere dried over Na₂SO₄ and concentrated. The residue was purified bychromatography on silica gel (15% EtOAc in hexanes) to afford{1S-[4R′-(4-fluoro-benzyl)-5-oxo-tetrahydrofuran-2S′-yl]-3-methylbutyl}-carbamicacid tert-butyl ester as a white foam (0.426 g, 31%): R_(f)=0.75 (25%EtOAc in hexanes); IR (cm⁻¹) 1765, 1702, 1510, 1186; ¹H NGR (CDCl₃) δ0.89 (d, 3H, J=6.6), 0.91 (d, 3H, J=6.6), 1.23-1.33 (m, 2H), 1.40 (s,9H), 1.45-1.52 (m, 1H), 1.96-2.05 (m, 1H), 2.22-2.32 (m, 1H), 2.77 (dd,1H, J=13.5, 8.7), 2.87-2.97 (m, 1H), 3.10 (dd, 1H, J=13.5, 4.2),3.74-3.82 (m, 1H), 4.25 (t, 1H, J=6.9), 4.34 (d, 1H, J=6.9), 6.95-7.01(m, 2H), 7.11-7.16 (m, 2H); Anal. (C₂₁H₃₀FNO₄) C, H, N.

[0382] Preparation of IntermediateEthyl-3-[BOC-L-LeuΨ[COCH₂]-L-(p-F)-Phe-L-(Tr-Gln)]-E-Propenoate.

[0383] To a solution of{1S-[4R′-(4-fluoro-benzyl)-5-oxo-tetrahydrofuran-2S′-yl]-3-methylbutyl}-carbamicacid tert-butyl ester (0.40 g, 1.06 mmol, 1 equiv) in1,2-dimethoxyethane (8 mL) was added LiOH (1.0 M solution in H₂O, 5.28mL, 5.28 mmol, 5.0 equiv). The reaction mixture was stirred at 23° C.for 20 min, then quenched with 0.5 M HCl (100 mL), and extracted with a1:1 mixture of EtOAc in hexanes (2×100 mL). The combined organic layerswere dried over Na₂SO₄ and concentrated. The residue was dissolved in a1:1 mixture of CH₂Cl₂ and CH₃CN (100 mL), and 4 Å molecular sieves (0.70g), 4-methylmorpholine N-oxide (0.248 g, 2.12 mmol, 2.0 equiv), andtetrapropylammonium perruthenate (0.037 g, 0.106 mmol, 0.1 equiv) wereadded sequentially. The reaction mixture was stirred at 23° C. for 1 hand filtered through celite. The filtrate was concentrated to give abrown oil which was dissolved in CH₂Cl₂ (15 mL). Crudeethyl-3-[H₂N-L-(Tr-Gln)]-E-propenoate•HCl (see preparation ofethyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-propenoate, 1.27 mmol,1.2 equiv), hydroxybenzotriazole hydrate (0.1 86g, 1.38 mmol, 1.3equiv), 4-methylmorpholine (0.466 mL, 4.24 mmol, 4.0 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.265 g,1.38 mmol, 1.3 equiv) were added sequentially. The reaction mixture wasstirred overnight, then poured into H₂O (50 mL), and extracted withCH₂Cl₂ (2×50 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated. The residue was purified by flash chromatography (40%EtOAc in hexanes) to provideethyl-3-[BOC-L-LeuΨ[COCH₂]-L-(p-F)-Phe-L-(Tr-Gln)]-E-propenoate as awhite foam: mp: 174-176° C.; R_(f)=0.56 (50% EtOAc in hexanes); IR(cm⁻¹) 1706, 1662, 1509; ¹H NGR (CDCl₃) (mixture of rotamers) δ 0.87(dd, J=6.9, 2.4), 1.02-1.12 (m), 1.27 (t, J=7.2), 1.39 (s), 1.96-2.02(m), 2.37 (t, J=7.2), 2.53 (d, J=12.3), 2.65 (dd, J=12.3, 5.7),2.76-2.99 (m), 3.62-3.68 (m), 4.17 (q, J=7.2), 4.47 (m), 4.60 (d,J=7.5), 5.39 (dd, J=15.9, 1.5), 5.97 (d, J=8.7), 6.61 (dd, J=15.3, 5.1),6.96 (t, J=8.4), 7.06-7.16 (m), 7.19-7.30 (m); Anal. (C₄₉H₅₈FN₅O₇) C, H,N.

[0384] Preparation of IntermediateEthyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-Propenoate.

[0385] Ethyl-3-[BOC-L-LeuΨ[COCH₂]-L-(p-F-Phe)-L-(Tr-Gln)]-E-propenoate(0.415 g, 0.50 mmol, 1 equiv) was dissolved in 1,4-dioxane (6 mL). Asolution of HCl in 1,4-dioxane (4.0 M, 6 mL) was added dropwise. Thereaction mixture was stirred for 2 h at 23° C., and then the solvent wasevaporated to provide the amine salt as a white foam. The crude aminesalt was dissolved in dry CH₂Cl₂ (10 mL) and cooled to 0° C.4-Methylmorpholine (0.166 mL, 1.51 mmol, 3.0 equiv) and cyclopentylchlorothiolformate (0.123 mL, 0.75 mmol, 1.5 equiv) were addedsequentially. The reaction mixture was stirred at 0° C. for 30 min,poured into H₂O (50 mL), and extracted with a 1:1 mixture of EtOAc andhexanes (3×50 mL). The combined organic layers were dried over Na₂SO₄and concentrated. The residue was purified by flash chromatography (50%EtOAc in hexanes) to provideethyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoateas a white foam (0.347 g, 83%): R_(f)=0.43 (50% EtOAc in hexanes); IR(cm⁻¹) 1716, 1651, 1510; ¹H NGR (CDCl₃) (mixture of rotamer) δ 0.87 (d,J=6.6), 1.06-1.16 (m), 1.29 (t, J=7.2), 1.53-1.60 (m), 1.67-1.69 (m),2.02-2.06 (m), 2.35 (t, J=7.2), 2.54 (d, J=15.3), 2.63-2.69 (m),2.78-2.97 (m), 3.57-3.68 (m), 4.17 (q, J=7.2), 4.38-4.49 (m), 4.42 (dd,J=18.0, 14.1), 6.05 (d, J=8.1), 6.62 (dd, J=15.6, 4.8), 6.94-7.00 (m),7.05-7.12 (m), 7.23-7.31 (m); Anal. (C₅₀H₅₈FN₃O₆S) C, H, N.

[0386] Preparation of ProductEthyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-F)Phe-L-Gln]-E-Propenoate.

[0387]Ethyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.285 g) was dissolved in CH₂Cl₂ (4 mL). Trifluoroacetic acid (4 mL)and triisopropylsilane (0.077 mL) were added sequentially to give abright yellow solution. After stirring for 30 min, no yellow colorremained. The solvents were evaporated to provide a white solid whichwas triturated with Et₂O (8 mL) and filtered to giveethyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-F)Phe-L-Gln]-E-propenoate aswhite solid (0.134 g, 65%): mp 179-180° C.; IR (cm⁻¹): 1718, 1656, 1511;¹H NGR (DMSO-d₆) δ 0.83 (d, 3H, J=6.0), 0.85 (d, 3H, J=6.0), 1.21 (t,3H, J=7.2), 1.30-1.44 (m, 4H), 1.53-1.70 (m, 7H), 1.97-2.05 (m, 4H),2.43-2.60 (m, 2H), 2.67-2.80 (m, 2H), 2.92-2.96 (m, 1H), 3.02-3.56 (m,2H), 4.09 (q, 2H, J=7.2), 4.17-4.19 (m, 1H), 4.31 (m, 1H), 5.42 (d, 1H,J=15.3), 6.62 (dd, 1H, J=15.3, 5.1), 6.75 (s, br, 1H), 7.00-7.06 (m,2H), 7.14-7.19 (m, 2H), 8.01 (d, 1H, J=8.1), 8.37 (d, 1H, J=7.8); Anal.(C₃₁H₄₄FN₃O₆S) C, H, N.

Example 20

[0388] Preparation of Compound 20:Ethyl-3-(CyPentylOCO-L-ValΨ[COCH₂]-L-p-CH₃)Phe-L-Gln)-E-Propenoate.

[0389] Preparation of IntermediateEthyl-3-[CyPentylOCO-L-VaIΨ[COCH₂]-L-p-CH₃)Phe-L-(Tr-Gln)]-E-Propenoate.

[0390] HCl (3 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution ofethyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate (0.311g, 0.388 mmol) in 1,4-dioxane (3 mL). The reaction solution was stirredat 23° C. for 2 hours and then concentrated. The residue was dissolvedin dry CH₂Cl₂ (6 mL). N,N-diisopropylethylamine (0.203 mL, 1.17 mmol, 3equiv) and cyclopentyl chloroformate (0.098 mL, 1.7 equiv) were addedsequentially. The reaction solution was stirred 3 hours at 23° C. andthen partitioned between brine (15 mL) and CH₂Cl₂ (3×30 mL). Thecombined organic phases were dried over Na₂SO₄, concentrated, and theresidue was chromatographed on silica gel (40% EtOAc in hexanes) toaffordethyl-3-[CyPentylOCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate(0.189 g, 60%) as a white foam: R_(f)=0.22 (40% EtOAc in hexanes); IR(cm⁻¹) 3316, 1712, 1667; ¹H NGR (CDCl₃) δ 0.65 (d, 3H, J=6.8), 0.92 (d,3H, J=6.8), 1.28 (t, 3H, J=7.2), 1.50-1.87 (m, 10H), 1.91-2.05 (m, 2H),2.29-2.38 (m, 1H), 2.31 (s, 3H), 2.51 (d, 1H, J=16.8), 2.60-2.69 (m,1H), 2.72-2.89 (m, 2H), 3.00 (dd, 1H, J=17.3, 9.8), 4.08-4.22 (m, 3H),4.42-4.53 (m, 1H), 4.88 (d, 1H, J=8.1), 4.95-5.02 (m, 1H), 5.49 (dd, 1H,J=15.8, 1.6), 5.88 (d, 1H, J=8.4), 6.60 (dd, 1H, J=15.8, 5.1), 7.00 (d,2H, J=7.9), 7.09 (d, 2H, J=7.9), 7.17-7.31 (m, 16H); Anal. (C₅₀H₅₉N₃O₇)C, H, N.

Example 21

[0391] Preparation of Compound 21:Ethyl-3-(CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-Propenoate.

[0392] Preparation of Intermediate{1S-[4R′-(4-Methyl-benzyl)-5-oxo-tetrahydrofuran-2S′-yl]-3-methylbutyl}-carbamicAcid tert-Butyl Ester.

[0393] n-Butyllithium (8.43 mL of a 1.6 M solution in hexanes, 13.48mmol, 2.0 equiv) was added to a solution of(3S-methyl-1R-oxiranyl-butyl)-carbamic acid tert-butyl ester (1.55 g,6.74 mmol, 1.0 equiv) and 3-p-tolyl-propionic acidcis-1S-amino-2R-indanol-acetonide amide (2.39 g, 7.03 mmol, 1 equiv) inTHF (100 mL) at −78° C. The resulting yellow solution was stirred at−78° C. for 5 min and then warmed to 0° C. for 1 h. The reaction wasquenched with 0.5 M HCl (100 mL) and extracted with a 1:1 mixture ofEtOAc and hexanes (2×100 mL). The combined organic layers were washedwith brine (150 mL), dried over Na₂SO₄, and concentrated. The residuewas purified by chromatography on silica gel (20% EtOAc in hexanes) toafford the coupling product as a white foam (2.07 g, 54%): R_(f)=0.30(25% EtOAc in hexanes); IR (cm⁻¹) 3415, 2955, 1687, 1612, 1355, 1166; ¹HNGR (CDCl₃) (mixture of diastereomers) δ 0.91 (d, J=6.3), 0.92 (d,J=6.9), 1.34 (s), 1.40 (s), 1.59 (s), 1.65-1.73 (m), 1.82-1.84 (m), 2.31(s), 2.36 (s), 2.76 (dd, J=12.9, 6.0), 3.06 (s), 3.20 (m), 3.34-3.47(m), 3.68 (m), 4.58 (d, J=9.3), 4.83 (s, br), 5.66 (m), 6.26 (d, J=7.5),6.85-6.90 (m), 7.09-7.24 (m).

[0394] This material was dissolved in a 5:1 mixture of toluene andCH₂Cl₂ (120 mL) and was treated with p-toluenesulfonic acid monohydrate(0.697 g, 3.67 mmol, 1.0 equiv). The reaction mixture was stirred at 23°C. for 14 h and then filtered through a medium frit. The clear filtratewas poured into half-saturated NaHCO₃ (100 mL) and extracted with a 1:1mixture of EtOAc in hexanes (2×100 mL). The organic layers were driedover Na₂SO₄ and concentrated. The residue was purified by flashchromatography on silica gel (10% EtOAc in hexanes) to afford{1S-[4R′-(4-methyl-benzyl)-5-oxo-tetrahydrofuran-2S′-yl]-3-methylbutyl}-carbamicacid tert-butyl ester as a white foam (0.572 g, 32%): IR (cm⁻¹) 1765,1707, 1167; ¹H NGR (CDCl₃) δ 0.89 (d, 3H, J=6.3), 0.90 (d, 3H, J=6.6)1.19-1.33 (m, 2H), 1.42 (s, 9H), 1.43-1.54 (m, 1H), 1.96-2.06 (m, 1H),2.18-2.27 (m, 1H), 2.32 (s, 3H), 2.75 (dd, 1H, J=13.5, 9.0), 2.88-2.98(m, 1H), 3.10 (dd, 1H, J=13.5, 4.2), 3.72-3.80 (m, 1H), 4.24 (t, 1H,J=6.3), 4.34 (d, 1H, J=9.9), 7.04-7.12 (m, 4H); Anal. (C₂₂H₃₃NO₄) C, H,N.

[0395] Preparation of IntermediateEthyl-3-[BOC-L-LeuΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)-E-Propenoate.

[0396] This material was prepared from{1S-[4R′-(4-methyl-benzyl)-5-oxo-tetrahydrofuran-2S′-yl]-3-methylbutyl}-carbamicacid tert-butyl ester (0.572 g, 1.52 mmol) as described previously forthe formation ofethyl-3-[BOC-L-LeuΨ[COCH₂]-L-(p-F)-Phe-L-(Tr-Gln)]-E-propenoate (Example19) to giveethyl-3-[BOC-L-LeuΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)-E-propenoate as awhite foam (0.785 g, 63%): R_(f)=0.70 (50% EtOAc in hexanes); IR (cm⁻¹)1709, 1664, 1170; ¹H NGR (CDCl₃) (mixture of rotamer) δ 0.85 (d, J=6.3),0.98-1.08 (m), 1.28 (t, J=7.2), 1.39 (s), 1.52-1.62 (m), 1.97-2.05 (m),2.31 (s), 2.36 (t, J=6.9), 2.52 (d, J=16.5), 2.64-2.67 (m), 2.83-1.97(m), 4.17 (q, J=7.2), 4.48 (m), 4.58 (d, J=7.2), 5.35 (dd, J=15.9, 1.5),5.87 (d, J=8.4), 6.61 (dd, J=15.9, 5.1), 7.02-7.11 (m), 7.20-7.30 (m);Anal. (C₅₀H₆₁N₃O₇.0.1H₂O) C, H, N.

[0397] Preparation of IntermediateEthyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-p-CH₃)Phe-L-(Tr-Gln]-E-Propenoate.

[0398] Ethyl-3-[BOC-L-LeuΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)-E-propenoate(0.523 g, 0.64 mmol) was deprotected and coupled with cyclopentylchlorothiolformate (0.158 mL, 0.96 mmol) as described previously for theformation ofethyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)-E-propenoate(Example 19) to giveethyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)-E-propenoateas a white foam (0.301 g, 56%): IR (cm⁻¹) 1716, 1651, 1518; ¹H NGR(CDCl₃) (mixture of rotamers) δ 0.85 (d, J=6.3), 1.02-1.12 (m,), 1.29(t, J=7.2), 1.52-1.62 (m), 1.67-1.68 (m), 1.98-2.10 (m), 2.31-2.36 (m),2.53 (d, J=15.0), 2.63-2.68 (m), 2.76-2.95 (m), 3.57-3.66 (m), 4.16 (q,J=7.2), 4.38 (m), 4.48-4.52 (m), 5.35 (d, J=7.5), 5.53 (dd, J=15.9,1.5), 5.94 (d, J=8.2), 6.62 (dd, J=15.9, 4.8), 6.99-7.12 (m), 7.21-7.31(m); Anal. (C₅₁H₆₁N₃O₆S.0.25H₂O) C, H, N.

[0399] Preparation of ProductEthyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln]-E-Propenoate.

[0400]Ethyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)-E-propenoate(0.272 g, 0.32 mmol) was deprotected using the procedure described forthe formation ofethyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-F)Phe-L-Gln]-E-propenoate(Example 19) to giveethyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln]-E-propenoate aswhite solid (0.106 g, 55%): mp 173-174° C.; IR (cm⁻¹) 1719, 1655, 1522,1198; ¹H NGR (DMSO-d₆) δ 0.83 (d, 3H, J=6.0), 0.85 (d, 3H, J=6.0), 1.21(t, 3H, J=7.2), 1.30-1.44 (m, 4H), 1.53-1.70 (m, 7H), 1.97-2.05 (m, 4H),2.43-2.60 (m, 2H), 2.67-2.80 (m, 2H), 2.92-2.96 (m, 1H), 3.02-3.56 (m,2H), 4.09 (q, 2H, J=7.2), 4.17-4.19 (m, 1H), 4.31 (m, 1H), 5.42 (d, 1H,J=15.3), 6.62 (dd, 1H, J=15.3, 5.1), 6.75 (s, br, 1H), 7.00-7.06 (m,2H), 7.14-7.19 (m, 2H), 8.01 (d, 1H, J=8.1), 8.37 (d, 1H, J=7.8); Anal.(C₃₁H44FN₃O₆S) C, H, N.

Example 22

[0401] Preparation of Compound 22:Ethyl-3-(CyPentylSCO-L-tert-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate.

[0402] Preparation of Intermediate CBZ-L-tert-Leucine.

[0403] To a solution of L-tert-leucine (5.12 g, 39 mmol, 1 equiv) in1,4-dioxane (80 mL) and 1 M NaOH (39 mL, 39 mmol, 1.0 equiv) at 0° C.was added benzyl chloroformate (6.24 mL, 43.68 mmol, 1.12 equiv). Thereaction mixture was stirred at 23° C. overnight, then concentrated,poured into 1 M HCl (80 mL), and extracted with CH₂Cl₂ (2×100 mL). Theorganic layers were dried over Na₂SO₄ and concentrated to give crudeCBZ-L-tert-leucine (20.7 g) as a clear oil: IR (cm⁻¹) 3336, 1715, 1521,1232; ¹H NGR (CDCl₃) δ 1.02 (s, 9H), 4.21 (d, 1H, J=9.9), 5.11 (s, 2H),5.35 (d, 1H, J=9.0), 7.34-7.37 (m, 5H).

[0404] Preparation of Intermediate4-Benzyloxycarbonylamino-5,5-dimethyl-3-oxo-hexanoic Acid tert-ButylEster.

[0405] To a solution of CBZ-L-tert-leucine (20.53 g, 77.4 mmol, 1 equiv)in THF (150 mL) was added 1,1′-carbonyldiimidazole (13.81 g, 85.14 mmol,1.1 equiv) at 23° C. The resulting solution was stirred at 23° C. for 1h. In a separate flask, n-butyllithium (101.59 mL of a 1.6 M solution inhexanes, 162.54 mmol, 2.1 equiv) was added to a solution ofdiisopropylamine (22.78 mL, 162.54 mmol, 2.1 equiv) in THF (100 mL) at−78° C. The reaction mixture was stirred for 15 min at −78° C., warmedto 0° C. for 5 min, then cooled back to −78° C. A solution of tert-butylacetate (21.9 mL, 162.54 mmol, 2.1 equiv) in THF (10 mL) was added viacannula, and the resulting mixture was stirred at −78° C. for 10 min.The above imidazole solution was then added dropwise to the lithiumenolate at −78° C. The resulting mixture was stirred at −78° C. for 1 h,quenched with 1 M HCl (100 mL), and extracted with EtOAc (2×100 mL). Thecombined organic layers were washed with brine (150 mL), dried overNa₂SO₄, and concentrated. The residue was purified by flashchromatography on silica gel (10% EtOAc in hexanes) to afford4-benzyloxycarbonylamino-5,5-dimethyl-3-oxo-hexanoic acid tert-butylester as a pale yellow oil (12.06 g, 44%): IR (cm⁻¹) 1717, 1508, 1265,739; ¹H NGR (CDCl₃) δ 1.09 (s, 9H), 1.44 (s, 9H), 3.50 (s, 2H), 4.29 (d,1H, J=9.3), 5.03 (s, 2H), 5.35-5.42 (m, 1H), 7.34 (s, 5H).

[0406] Preparation of Intermediate CBZ-L-tert-LeuΨ[COCH₂]-L-Phe-OMe.

[0407] To a stirred solution of (R)-2-hydroxy-3-phenyl-propionic acidmethyl ester (2.53 g, 14.06 mmol, 3.1 equiv) in CH₂Cl₂ (30 mL) at 0° C.was added trifluoromethanesulfonic anhydride (2.50 mL, 14.8 mmol, 3.3equiv) and 2,6-lutidine (1.72 mL, 14.8 mmol, 3.3 equiv) slowly. Theresulting pink solution was stirred at 0° C. for 30 min, then pouredinto 0.5 M HCl (100 mL), and extracted with a 1:1 mixture of EtOAc inhexanes (2×100 mL). The combined organic layers were dried over Na₂SO₄,concentrated, and utilized in the next step below.

[0408] A solution of4-benzyloxycarbonylamino-5,5-dimethyl-3-oxo-hexanoic acid tert-butylester (1.64 g, 4.51 mmol, 1 equiv) in THF (100 mL) was added dropwise toa stirred suspension of NaH (0.190 g of a 60% dispersion in mineral oil,4.74 mmol, 1.05 equiv) in THF (100 mL) at 0° C. After stirring for 10min, a solution of crude (R)-2-triflyoxy-3-phenyl-propionic acid methylester (prepared above) in CH₂Cl₂ (10 mL) was added dropwise. Theresulting mixture was stirred at 23° C. for 24 h, then quenched with 1 MHCl (50 mL), and extracted with EtOAc (3×50 mL). The combined organiclayers were washed with brine (100 mL), dried over Na₂SO₄, andconcentrated to provide a pale yellow oil. Without further purification,the above oil was dissolved in CH₂Cl₂ (10 mL), treated withtrifluoroacetic acid (2 mL), and then maintained at 23° C. for 24 h.After dilution with CH₂Cl₂ (50 mL), the resulting solution was washedwith saturated NaHCO₃ (50 mL) and brine (50 mL). The organic layer wasdried over Na₂SO₄ and concentrated. The residue was purified by flashchromatography on silica gel (10% EtOAc in hexanes) to affordCBZ-L-tert-LeuΨ[COCH₂]-L-Phe-OMe as a pale yellow oil (1.01 g, 54%):R_(f)=0.41 (25% EtOAc in hexanes); IR (cm⁻¹) 1711, 1514, 1233; ¹H NGR(CDCl₃) δ 0.97 (s, 9H), 2.58-2.76 (m, 2H), 2.96-3.17 (m, 3H), 3.62 (s,3H), 4.17 (d, 1H, J=8.1), 5.06-5.10 (s, 2H), 5.32 (d, 1H, J=8.6),7.12-7.36 (m, 10 H); Anal. (C₂₅H₃₁NO₅.0.25 H₂O) C, H, N.

[0409] BOC-L-tert-LeuΨ[COCH₂]-L-Phe-OMe.

[0410] 10% Pd on C (0.110 g) was added to a solution ofCBZ-L-tert-LeuΨ[COCH₂]-L-Phe-OMe (0.513 g, 1.33 mmol, 1 equiv) anddi-tert-butyl dicarbonate (0.378 g, 1.73 mmol, 1.3 equiv) in CH₃OH at23° C. The reaction mixture was stirred at 23° C. under an H₂ atmosphere(balloon) overnight. The mixture was filtered through celite, and thefiltrate was concentrated. The residue was purified by flashchromatography on silica gel (10% EtOAc in hexanes) to affordBOC-L-tert-LeuΨ[COCH₂]-L-Phe-OMe as white solid (0.366 g, 70%):mp=98-99° C.; R_(f)=0.54 (25% EtOAc in hexanes); IR (cm⁻¹) 1707, 1497,1367, 1236, 1168; ¹H NGR (CDCl₃) δ 0.97 (s, 9H), 1.40 (s, 9H), 2.60-2.78(m, 2H), 2.95-3.19 (m, 3H), 3.63 (s, 3H), 4.07-4.11 (m, 2H), 5.07 (d,1H, J=9.3), 7.13-7.32 (m 5H); Anal. (C₂₅H₃₁NO₅) C, H, N.

[0411] Preparation of Intermediate CBZ-L-tert-LeuΨ[COCH₂]-L-Phe-OH.

[0412] 2 M NaOH (3.35 mL, 6.7 mmol, 8.0 equiv) was added to a solutionof BOC-L-tert-LeuΨ[COCH₂]-L-Phe-OMe (0.328 g, 0.84 mmol, 1 equiv) inCH₃OH (6 mL) at 0° C. over 10 min. The reaction mixture was stirred at0° C. for 2 h, then poured into 10% KHSO₄ (80 mL), and extracted withCH₂Cl₂ (2×100 mL). The organic layers were dried over Na₂SO₄ andconcentrated to give BOC-L-tert-LeuΨ[COCH₂]-L-Phe-OH as a white solid(0.315 g, 99%) which was used without further purification: IR (cm⁻¹)2960, 1710, 1498, 1368, 1167; ¹ H NGR (CDCl₃) δ 0.94 (s, 9H), 1.39 (s,9H), 2.60-2.80 (m, 2H), 2.95-3.16 (m, 3H), 4.08 (d, 2H, J=9.3), 5.09 (d,1H, J=9.6), 7.12-7.31 (m 5H).

[0413] Preparation of IntermediateEthyl-3-[BOC-L-tert-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenoate.

[0414] Ethyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (0.523 g, 0.64 mmol) wasdeprotected and coupled with BOC-L-tert-LeuΨ[COCH₂]-L-Phe-OH (0.315 g,0.84 mmol) as described for the formation ofethyl-3-[BOC-L-LeuΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate (Example19) to giveethyl-3-[BOC-L-tert-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate as awhite foam (0.474 g, 70%): R_(f)=0.58 (50% EtOAc in hexanes); IR (cm⁻¹)1702, 1669, 1494, 1169; ¹H NGR (CDCl₃) (mixture of rotamers) δ 0.85 (s),1.30 (t, J=7.2), 1.41 (s), 1.56-1.65 (m), 1.95-2.02 (m), 2.22-2.42 (m),2.62-2.88 (m), 3.09-3.18 (m), 4.00 (d, J=8.7), 4.17 (t, J=7.2),4.46-4.51 (m), 4.93 (d, J=8.7), 5.37 (d, J=15.9), 5.69 (d, J=9.3), 6.54(dd, J=15.9, 4.8), 7.19-7.15 (m), 7.18-7.31 (m); Anal. (C₄₉H₅₉N₃O₇) C,H, N.

[0415] Preparation of IntermediateEthyl-3-[CyPentylSCO-L-tert-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-Propenoate.

[0416] Ethyl-3-[BOC-L-tert-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.441 g, 0.55 mmol) was deprotected and coupled with cyclopentylchlorothiolformate (0.135 mL, 0.82 mmol) as described previously for theformation ofethyl-3-[CyPentylSCO-L-LeuΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(Example 19) to giveethyl-3-[CyPentylSCO-L-tert-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoateas a white foam (0.347 g, 76%): IR (cm⁻¹) 1718, 1656, 1493, 1186; ¹H NGR(CDCl₃) (mixture of rotamers) δ 0.86(s), 1.27 (t, J=7.2), 1.56-1.68 (m),1.95-2.12 (m), 2.22-2.39 (m), 2.60-2.74 (m), 2.84-2.90 (m), 3.05-3.14(m), 3.59-3.64 (m), 4.16 (q, J=7.2), 4.31 (d, J=8.4), 4.48 (m), 5.41(dd, J=15.9, 1.8), 5.67 (d, J=8.7), 5.82 (d, J=9.0), 6.56 (dd, J=15.6,5.1), 7.19-7.31 (m); Anal. (C₅₀H₅₉N₃O₆) C, H, N.

[0417] Preparation of ProductEthyl-3-(CyPentylSCO-L-tert-LeuΨ[COCH₂]-L-Phe-L-Gln)-E-Propenoate.

[0418]Ethyl-3-[CyPentylSCO-L-tert-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.318 g, 0.38 mmol) was deprotected using the procedure described forthe formation ofethyl-3-(CyPentylSCO-L-LeuΨ[COCH₂]-L-(P-F)Phe-L-Gln)-E-propenoate(Example 19) to giveethyl-3-[CyPentylSCO-L-tert-LeuΨ[COCH₂]-L-Phe-L-Gln]-E-propenoate aswhite solid (0.204 g, 91%): mp 65-68° C.; IR (cm⁻¹) 1715, 1652, 1520,1193; ¹H NGR (CDCl₃) (mixture of rotamers) δ 0.96 (s), 1.31 (t, J=7.2),1.50-1.73 (m), 1.96-2.13 (m), 2.23 (t, J=7.5), 2.68-2.79 (m), 2.84-2.95(m), 3.11-3.21 (m), 3.59-3.69 (m), 4.18 (q, J=7.2), 4.36 (d, J=8.1),4.52-4.59 (m), 5.37 (s, br), 5.42 (dd, 1H, J=15.9, 1.5), 5.80 (d,J=9.0), 5.90 (d, J=8.4), 6.46 (s, br), 6.61 (dd, 1H, J=15.9, 5.1),7.18-7.30 (m); Anal. (C₃₁H₄₅N₃O₆S) C, H, N.

[0419] Preparation of ProductEthyl-3-(CyPentylSCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-Propenoate.

[0420]Ethyl-3-[CyPentylOCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate(0.160 g, 0.197 mmol, 1 equiv) was dissolved in dry CH₂Cl₂ (8 mL).Triisopropylsilane (0.121 mL, 0.591 mmol, 3 equiv) and trifluoroaceticacid (4 mL) were added sequentially producing a bright yellow solution.This solution was stirred for 40 minutes and then concentrated. Theresidue was stirred in Et₂O (8 mL), and the solid was collected byfiltration, washed with Et₂O (2×4 mL), and then dried under vacuum togive ethyl-3-(CyPentylOCO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-propenoate(0.094 g, 84%) as a white solid: mp =206-207° C. (dec); R_(f)=0.38 (10%CH₃OH in CH₂Cl₂); IR (cm⁻¹) 3413, 3307, 3213, 1708, 1660; ¹H NGR(DMSO-d₆) δ 0.72 (d, 3H, J=6.8), 0.81 (d, 3H, J=6.5), 1.20 (t, 3H,J=7.0), 1.46-1.82 (m, 10H), 1.97-2.10 (m, 3H), 2.23 (s, 3H), 2.38-2.54(m, 1H), 2.65-2.84 (m, 2H), 2.86-2.97 (m, 1H), 3.81-3.88 (m, 1H),4.03-4.18 (m, 2H), 4.26-4.38 (m, 1H), 4.85-4.94 (m, 1H), 5.55 (d, 1H,J=15.9), 6.65 (dd, 1H, J=15.9, 5.3), 6.73 (s, 1H), 7.03 (s, 4H), 7.14(s, 1H), 7.27 (d, 1H, J=8.1), 8.02 (d, 1H, J=8.4); Anal. (C₃₁H₄₅N₃O₇) C,H, N.

Example 23

[0421] Preparation of Compound 23:Ethyl-3-(CyPentylCH₂CO-L-ValΨ[COCH₂]-L-P-CH₃)Phe-L-Gln)-E-Propenoate.

[0422] Preparation of IntermediateEthyl-3-[CyPentylCH₂CO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-Propenoate.

[0423] HCl (3 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution ofethyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate (0.315g, 0.393 mmol, 1 equiv) in 1,4-dioxane (3 mL). The reaction solution wasstirred at 23° C. for 1.67 hours and then was concentrated to providecrudeethyl-3-[H₂N-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoatec•HClwhich was set aside.

[0424] Dicyclohexylcarbodiimide (0.162 g, 0.785 mmol, 2 equiv) was addedto a solution of cyclopentyl acetic acid (0.197 mL, 1.57 mmol, 4 equiv)in Et₂O (10 mL). The reaction mixture was stirred for 1.5 hours, andthen the white precipitate was removed by filtration. The filtrate wasconcentrated, then dissolved in dry CH₂Cl₂ (5 mL), and added to asolution of crudeethyl-3-[H₂N-L-ValΨ[COCH₂]-L-p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate-HCl(from above) and 4-methylmorpholine (0.086 mL, 0.782 mmol, 2 equiv) indry CH₂Cl₂ (3 mL). The reaction mixture was stirred for 2.5 hours andconcentrated. The residue was chromatographed on silica gel (gradientelution 40→50% EtOAc in hexanes) to affordethyl-3-[CyPentylCH₂CO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate(0.202 g, 63%) as a white foam: R_(f)=0.65 (10% CH₃OH in CHCl₃); IR(cm⁻¹) 3296, 1716, 1650; ¹H NGR (CDCl₃) δ 0.71 (d, 3H, J=6.8), 0.92 (d,3H, J=6.8), 1.02-1.16 (m, 2H), 1.29 (t, 3H, J=7.2), 1.47-1.82 (m, 8H),1.89-2.16 (m, 5H), 2.30-2.36 (m, 1H), 2.31 (s, 3H), 2.46-2.69 (m, 2H),2.74-2.89 (m, 2H), 2.95-3.08 (m, 1H), 4.12-4.22 (m, 2H), 4.41-4.53 (m,2H), 5.52 (dd, 1H, J=15.6, 1.6), 5.73 (d, 1H, J=8.1), 6.00 (d, 1H,J=8.1), 6.61 (dd, 1H, J=15.6, 5.1), 6.99 (d, 2H, J=7.9), 7.08 (d, 2H,J=7.9), 7.17-7.30 (m, 16H); Anal. (C₅₁H₆₁N₃O₆.0.5 H₂O) C, H, N.

[0425] Preparation of ProductEthyl-3-(CyPentylCH₂CO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-Propenoate.

[0426]Ethyl-3-[CyPentylCH₂CO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-(Tr-Gln)]-E-propenoate(0.167 g, 0.206 mmol, 1 equiv) was dissolved in dry CH₂Cl₂ (8 mL).Triisopropylsilane (0.126 mL, 0.615 mmol, 3 equiv) and trifluoroaceticacid (4 mL) were added sequentially producing a bright yellow solution.This solution was stirred for 40 minutes and then concentrated. Theresidue was stirred in Et₂O (8 mL), and the solid was collected byfiltration, washed with Et₂O (2×4 mL), and then dried under vacuum togiveethyl-3-(CyPentylCH₂CO-L-ValΨ[COCH₂]-L-(p-CH₃)Phe-L-Gln)-E-propenoate(0.092 g, 79%) as a white solid: mp=253-255 ° C. (dec); R_(f)=0.42 (10%CH₃OH in CHCl₃); IR (cm⁻¹) 3401, 3284, 1713, 1649; ¹H NGR (DMSO-d₆) δ0.74 (d, 3H, J=6.8), 0.81 (d, 3H, J=6.5), 1.03-1.19 (m, 2H), 1.20 (t,3H, J=7.0), 1.40-1.75 (m, 9H), 2.00-2.15 (m, 6H), 2.23 (s, 3H), 2.42(dd, 1H, J=18.4, 4.7), 2.66-2.84 (m, 2H), 2.86-2.96 (m, 1H), 4.06-4.17(m, 3H), 4.27-4.37 (m, 1H), 5.57 (dd, 1H, J=15.7, 1.4), 6.66 (dd, 1H,J=15.7, 5.4), 6.73 (s, 1H), 7.03 (s, 4H), 7.16 (s, 1H), 7.92 (d, 1H,J=8.1), 8.01 (d, 1H, J=7.8); Anal. (C₃₂H₄₇N₃O₆) C, H, N.

Example 24

[0427] Preparation of Compound 24:Ethyl-3-(CyPentylSCO-L-CyhexΨ[COCH₂]-L-(p-F)Phe-L-Gln)-E-Propenoate.

[0428] Preparation of Intermediate trans-5-Cyclohexyl-pent-4-enoic Acid.

[0429] A solution of cyclohexane carboxaldehyde (11.22 g, 100 mmol, 1equiv) in THF (100 mL) was added dropwise via addition funnel to asolution of vinylmagnesium bromide (100 mL of a 1.0 M solution in THF,100 mmol, 1.0 equiv) in Et₂O (100 mL) at 0° C. After the addition wascompleted, the reaction mixture was stirred for 1 h at 0° C. and thenpartitioned between 0.5 M HCl (150 mL) and a 1:1 mixture of EtOAc andhexanes (2×150 mL). The combined organic layers were dried over Na₂SO₄and concentrated to afford a yellow oil. This material was combined(neat) with diethyl malonate (16.7 mL, 110 mmol, 1.1 equiv) and Ti(OEt)₄(2.10 mL, 10.0 mmol, 0.10 equiv) and was heated to 160° C. for 1 h(distilling out EtOH as it was formed). The reaction mixture was thenmaintained at 190° C. for 4 h and then cooled to 60° C. EtOH (50 mL) and6.0 M KOH (50 mL) were added sequentially, and the brown reactionmixture was refluxed for 3 h. After cooling to 23° C., the reactionmixture was filtered through a medium frit, and the filtrate waspartitioned between water (150 mL) and Et₂O (2×100 mL). The aqueouslayer was then acidified to pH=2 (as indicated by pH paper) withconcentrated HCl and extracted with a 1:1 mixture of EtOAc and hexanes(2×150 mL). The combined organic layers were dried over Na₂SO₄,concentrated, and the residue was distilled at reduced pressure toafford trans-5-cyclohexyl-pent-4-enoic acid (5.68 g, 31%) as a colorlessliquid: bp: 150-156° C. (1 torr); IR (cm⁻¹) 3001 (br), 2923, 1711; ¹HNGR (CDCl₃) δ 0.96-1.32 (m, 5H), 1.60-1.76 (m, 5H), 1.85-1.94 (m, 1H),2.27-2.44 (m, 4H), 5.31-5.48 (m, 2H); Anal. (C₁₁H₁₈O₂) C, H.

[0430] Preparation of Intermediate trans-5-Cyclohexyl-pent-4-enoic Acid(2R-Hydroxy-1R-methyl-2-phenyl-ethyl)-methyl Amide.

[0431] Oxalyl chloride (2.81 mL, 32.2 mmol, 1.05 equiv) was added to asolution of trans-5-cyclohexyl-pent-4-enoic acid (5.60 g, 30.7 mmol, 1equiv) and N,N-dimethylformamide (0.03 mL, 0.39 mmol, 0.013 equiv) inbenzene (100 mL) at 23° C. The reaction mixture was stirred at 23° C.for 2 h and then concentrated under reduced pressure. The resulting oilwas dissolved in THF (20 mL) and added via cannula to a solution of(1R,2R)-(−)-pseudoephedrine (4.61 g, 27.9 mmol, 0.91 equiv) andtriethylamine (5.06 mL, 36.3 mmol, 1.1 equiv) in THF (300 mL) at 0° C.The reaction mixture was stirred at 0° C. for 30 min and thenpartitioned between half-saturated NH₄Cl (150 mL) and a 1:1 mixture ofEtOAc and hexanes (2×150 mL). The combined organic layers were driedover Na₂SO₄, concentrated, and the residue purified by flash columnchromatography (50% EtOAc in hexanes) to affordtrans-5-cyclohexyl-pent-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (5.21 g, 57%) as awhite solid: mp=89-91° C.; R_(f)=0.33 (50% EtOAc in hexanes); IR (cm⁻¹)3380, 1621; ¹H NGR (CDCl₃, mixture of rotamers) δ 0.97-1.32 (m),1.54-1.74 (m), 1.86-1.93 (m), 2.24-2.58 (m), 2.81 (s), 2.91 (s),3.98-4.06 (m), 4.35-4.48 (m), 4.55-4.61 (m), 5.32-5.47 (m), 7.24-7.41(m); Anal. (C₂₁H₃₁NO₂) C, H, N.

[0432] Preparation of Intermediatetrans-5-Cyclohexyl-2S-(4-fluoro-benzyl)-pent-4-enoic Acid(2R-Hydroxy-1R-methyl-2-phenyl-ethyl)-methyl Amide.

[0433] n-Butyllithium (20.7 mL of a 1.6 M solution in hexanes, 33.1mmol, 2.1 equiv) was added to a suspension of anhydrous lithium chloride(4.68 g, 110 mmol, 7 equiv) and diisopropylamine (4.98 mL, 35.5 mmol,2.25 equiv) in THF (300 mL) at −78° C. The reaction mixture was stirredfor 20 min at −78° C., then maintained at 0° C. for 5 min, andsubsequently cooled again to −78° C. A solution oftrans-5-cyclohexyl-pent-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (5.20 g, 15.8 mmol, 1equiv) in THF (30 mL) was added via cannula, and the resulting solutionwas stirred at −78° C. for 1 h, maintained at 0° C. for 15 min, stirredat 23° C. for 5 min and then cooled again to 0° C. 4-Fluorobenzylbromide (2.95 mL, 23.7 mmol, 1.5 equiv) was added, and the reactionmixture was stirred at 0° C. for 30 min and then partitioned betweenhalf-saturated NH₄Cl (150 mL) and a 1:1 mixture of EtOAc and hexanes(2×150 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated. Purification of the residue by flash column chromatography(gradient elution 30→40% EtOAc in hexanes) providedtrans-5-cyclohexyl-2S-(4-fluoro-benzyl)-pent-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (6.29 g, 91%) as aviscous oil: R_(f)=0.57 (50% EtOAc in hexanes); IR (cm⁻¹) 3382, 1616; ¹HNGR (CDCl₃, mixture of rotamers) δ 0.83-1.27 (m), 1.51-1.70 (m),1.86-1.95 (m), 2.12-2.21 (m), 2.28-2.44 (m), 2.58 (s), 2.64-2.77 (m),2.82 (s), 2.85-2.92 (m), 4.00-4.05 (m), 4.37-4.52 (m), 5.22-5.52 (m),6.88-7.01 (m), 7.07-7.20 (m), 7.20-7.38 (m); Anal. (C28H36FNO₂.0.25H₂O)C, H, N.

[0434] Preparation of Intermediate5S-(R-Bromo-cyclohexyl-methyl)-3R-(4-fluoro-benzyl)-dihydrofuran-2-one.

[0435] N-Bromosuccinimide (2.65 g, 14.9 mmol, 1.1 equiv) was added insmall portions over 5 min to a solution oftrans-5-cyclohexyl-2S-(4-fluoro-benzyl)-pent-4-enoic acid(2R-hydroxy-1R-methyl-2-phenyl-ethyl)-methyl amide (6.20 g, 14.2 mmol, 1equiv) and glacial acetic acid (3.87 mL, 70.8 mmol, 5 equiv) in a 4:1mixture of THF and H₂O (250 mL) at 0° C. The resulting yellow solutionwas stirred for 15 min at 0° C., then warmed to 23° C., and subsequentlyrefluxed for 1 h. After cooling to 23° C., the reaction mixture waspartitioned between half-saturated NaHCO₃ (150 mL) and a 1:1 mixture ofEtOAc and hexanes (2×150 mL). The combined organic layers were driedover Na₂SO₄ and concentrated. Flash chromatographic purification of theresidue (5% EtOAc in hexanes) gave5S-(R-bromo-cyclohexyl-methyl)-3R-(4-fluoro-benzyl)-dihydrofuran-2-one(3.70 g, 71%) as a white solid: mp=72-75° C.; R_(f)=0.62 (30% EtOAc inhexanes); IR (cm⁻¹) 1774; ¹H NGR (CDCl₃) δ 1.11-1.38 (m, 6H), 1.52-1.77(m, 5H), 2.13-2.34 (m, 2H), 2.82 (dd, 1H, J=13.7, 8.4), 2.95-3.05 (m,1H), 3.12 (dd, 1H, J=13.7, 4.7), 3.89 (dd, 1H, J=8.4, 3.7), 4.43-4.51(m, 1H), 6.98-7.05 (m, 2H), 7.15-7.26 (m, 2H); Anal. (C₁₈H₂₂BrFO₂) C, H,N.

[0436] Preparation of Intermediate5S-(S-Azido-cyclohexyl-methyl)-3R-(4-fluoro-benzyl)-dihydro-furan-2-one.

[0437] A suspension of sodium azide (1.30 g, 20.0 mmol, 2 equiv) and5S-(R-bromo-cyclohexyl-methyl)-3R-(4-fluoro-benzyl)-dihydrofuran-2-one(3.70 g, 10.0 mmol, 1 equiv) in N,N-dimethylformamide (30 mL) was heatedat 50° C. for 18 h. The reaction mixture was cooled to 23° C. and thenpartitioned between water (150 mL) and a 1:1 mixture of EtOAc andhexanes (2×150 mL). The combined organic layers were dried over Na₂SO₄,concentrated, and the residue purified by flash column chromatography(10% EtOAc in hexanes) to give5S-(S-azido-cyclohexyl-methyl)-3R-(4-fluoro-benzyl)-dihydro-furan-2-one(1.76 g, 53%) as a colorless oil: R_(f)=0.33 (20% EtOAc in hexanes); IR(cm⁻¹) 2109, 1772; ¹H NGR (CDCl₃) δ 1.06-1.29 (m, 6H), 1.67-1.82 (m,5H), 2.02-2.21 (m, 2H), 2.79-2.86 (m, 1H), 2.92-2.95 (m, 1H), 3.05-3.17(m, 2H), 4.45-4.50 (m, 1H), 6.97-7.04 (m, 2H), 7.15-7.26 (m, 2H).

[0438] Preparation of Intermediate{S-Cyclohexyl-[4R-(4-fluoro-benzyl)-5-oxo-tetrahydro-furan-2S-yl]-methyl}-carbamicAcid tert-Butyl Ester.

[0439] A suspension of5S-(S-azido-cyclohexyl-methyl)-3R-(4-fluoro-benzyl)-dihydro-furan-2-one(1.76 g, 5.31 mmol, 1 equiv) and Pd/C (10%, 0.15 g) in CH₃OH (30 mL) wasstirred under a hydrogen atmosphere (balloon) for 3 h. The reactionmixture was filtered through celite, concentrated, and the residuedissolved in 1,4-dioxane (30 mL). N,N-diisopropylethylamine (1.85 mL,10.6 mmol, 2 equiv) and di-tert-butyl dicarbonate (1.74 g, 7.97 mmol,1.5 equiv) were added sequentially, and the resulting solution wasstirred at 23° C. for 1.5 h. The reaction mixture was then partitionedbetween water (150 mL) and a 1:1 mixture of EtOAc and hexanes (2×150mL). The combined organic layers were dried over Na₂SO₄ andconcentrated. Purification of the residue by flash column chromatography(15% EtOAc in hexanes) provided{S-cyclohexyl-[4R-(4-fluoro-benzyl)-5-oxo-tetrahydro-furan-2S-yl]-methyl}-carbamicacid tert-butyl ester (1.11 g, 52%) as a white foam: R_(f)=0.55 (30%EtOAc in hexanes); IR (cm⁻¹) 3338, 1766, 1699; ¹H NGR (CDCl₃) δ0.92-1.26 (m, 5H), 1.40 (s, 9H), 1.62-1.80 (m, 6H), 1.95-2.05 (m, 1H),2.17-2.27 (m, 1H), 2.79 (dd, 1H, J=13.5, 8.6), 2.88-2.98 (m, 1H), 3.09(dd, 1H, J=13.5, 4.5), 3.37-3.43 (m, 1H), 4.43 (d, 1H, J=10.0),4.48-4.52 (m, 1H), 6.96-7.01 (m, 2H), 7.12-7.27 (m, 2H); Anal.(C₂₁H₃₁NO₄) C, H,N.

[0440] Preparation of IntermediateEthyl-3-[BOC-L-CyhexΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-Propenoate.

[0441] Lithium hydroxide (7.0 mL of a 1 M aqueous solution, 7.0 mmol, 5equiv) was added to a solution of{S-cyclohexyl-[4R-(4-fluoro-benzyl)-5-oxo-tetrahydro-furan-2S-yl]-methyl}-carbamicacid tert-butyl ester (0.567 g, 1.40 mmol, 1 equiv) in DME (10 mL) at23° C. The resulting suspension was stirred at 23° C. for 30 min andthen partitioned between 0.5 M HCl (100 mL) and EtOAc (2×100 mL). Thecombined organic layers were dried over Na₂SO₄, concentrated, and theresidue dissolved in CH₂Cl₂ (30 mL). 4-Methylmorpholine N-oxide (0.328g, 2.80 mmol, 2 equiv), powdered 4 Å molecular sieves (0.60 g), andtetrapropylammonium perruthenate (0.049 g, 0.139 mmol, 0.1 equiv) wereadded sequentially. The resulting dark reaction mixture was stirred for2 h at 23° C. and then filtered through celite. The filtrate wasconcentrated under reduced pressure to provide a brown oil which wasdissolved in CH₂Cl₂ (40 mL). Crudeethyl-3-[H₂N-L-(Tr-Gln)]-E-propenoate•HCl (see preparation ofethyl-3-[CBZ-L-LeuΨ[COCH₂]-D/L-Phe-L-(Tr-Gln)]-E-propenoate, 1.68 mmol,1.2 equiv), 1-hydroxybenzotriazole hydrate (0.284 g, 2.10 mmol, 1.5equiv), 4-methylmorpholine (0.616 mL, 5.60 mmol, 4 equiv), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.403 g,2.10 mmol, 1.5 equiv) were added sequentially, and the reaction mixturewas stirred for 20 h at 23° C. and then partitioned between water (150mL) and a 1:1 mixture of EtOAc and hexanes (2×150 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated. Purification ofthe residue by flash column chromatography (40% EtOAc in hexanes)providedethyl-3-[BOC-L-CyhexΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate (0.379g, 32%) as awhite solid: mp=192-195° C.; R_(f=)0.50 (50% EtOAc inhexanes); IR (cm⁻¹) 3316, 1709, 1667; ¹H NGR (CDCl₃) δ 0.98-1.12 (m,5H), 1.29 (t, 3H, J=7.2), 1.40 (s, 9H), 1.43-1.74 (m, 6H), 1.96-2.02 (m,1H), 2.30-2.39 (m, 2H), 2.53 (d, 1H, J=17.1), 2.61-2.88 (m, 4H), 3.00(dd, 1H, J=17.6, 10.1), 4.01-4.06 (m, 1H), 4.17 (q, 2H, J=7.2), 4.45 (s,br, 1H), 4.80 (d, 1H, J=8.1), 5.35 (d, 1H, J=15.7), 5.90 (d, H, J=8.4),6.60 (dd, 1H, J=15.7, 5.0), 6.93-7.10 (m, 4H), 7.17-7.30 (m, 16H); Anal.(C₅₁H₆₀FN₃O₇) C, H, N.

[0442] Preparation of IntermediateEthyl-3-[CyPentylSCO-L-CyhexΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-Propenoate.

[0443] HCl (10 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution of ethyl-3-[BOC-L-LeuΨ[COCH₂]-L-Phe-L-(Tr-Gln)]-E-propenoate(0.379 g, 0.448 mmol, 1 equiv) in 1,4-dioxane (10 mL). The reactionmixture was stirred at 23° C. for 2 h and then concentrated. Theresulting oil was dissolved in CH₂Cl₂ (15 mL), cooled to 0° C., and4-methylmorpholine (0.123 mL, 1.12 mmol, 2.5 equiv) and cyclopentylchlorothiolformate (0.096 mL, 0.583 mmol, 1.3 equiv) were addedsequentially. The reaction mixture was stirred for 30 min at 0° C. andthen partitioned between water (100 mL) and a 1:1 mixture of EtOAc andhexanes (2×100 mL). The combined organic layers were dried over Na₂SO₄,concentrated, and the residue was chromatographed on silica gel (30%EtOAc in hexanes) to affordethyl-3-[CyPentylSCO-L-CyhexΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.161 g, 41%) as a white solid: mp=90-95° C.; R_(f)=0.51 (50% EtOAc inhexanes); IR (cm⁻¹) 3312, 1714, 1654; ¹H NGR (CDCl₃) δ 0.98-1.22 (m,6H), 1.29 (t, 3H, J=7.2), 1.56-1.70 (m, 11 H), 1.98-2.08 (m, 3H),2.29-2.37 (m, 2H), 2.51-3.30 (m, 6H), 3.58-3.65 (m, 1H), 4.17 (q, 2H,J=7.2), 4.33 (s, br, 1H), 4.45 (s, br, 1H), 5.37 (dd, 1H, J=15.7, 1.6),5.55 (d, 1H, J=8.1), 5.99 (d, 1H, J=8.4), 6.61 (dd, 1H, J=15.7, 4.7),6.93-7.10 (m, 4H), 7.16-7.31 (m, 16H); Anal. (C₅₂H₆₀FN₃O₆S) C, H, N.

[0444] Preparation of ProductEthyl-3-(CyPentylSCO-L-CyhexΨ[COCH₂]-L-(p-F)Phe-L-Gln)-E-Propenoate.

[0445] Triisopropylsilane (0.10 mL) and trifluoroacetic acid (6 mL) wereadded sequentially to a solution ofethyl-3-[CyPentylSCO-L-CyhexΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.150 g, 0.172 mmol) in CH₂Cl₂ (8 mL) producing a bright yellowsolution. The reaction mixture was stirred for 20 min at 23° C., thencarbon tetrachloride (4 mL) was added, and the mixture was concentratedunder reduced pressure. The residue was purified by flash columnchromatography (3% CH₃OH in CH₂Cl₂) to affordethyl-3-(CyPentylSCO-L-CyhexΨ[COCH₂]-L-(p-F)Phe-L-Gln)-E-propenoate(0.069 g, 63%) as a white foam: R_(f)=0.56 (10% CH₃OH in CH₂Cl₂); IR(cm⁻¹) 3281, 1716, 1637; ¹H NGR (DMSO-d₆) δ 0.93-1.14 (m, 6H), 1.21 (t,3H, J=7.2), 1.41-1.70 (m, 13H), 1.96-2.03 (m, 4H), 2.53-2.93 (m, 5H),3.46-3.56 (m, 1H), 3.97-4.15 (m, 3H), 4.30 (s, br, 1H), 5.41 (d, 1H,J=15.7), 6.61 (dd, 1H, J=15.7, 5.0), 6.74 (s, 1H), 7.00-7.19 (m, 5H),8.00 (d, 1H, J=8.4), 8.28 (d, 1H, J=7.8); Anal. (C₃₃H₄₆FN₃O₆S) C, H, N.

Example 25

[0446] Preparation of Compound 25:Ethyl-3-(CyPentylOCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-Gln)-E-Propenoate.

[0447] Preparation of IntermediateEthyl-3-[CyPentylOCO-L-VaIΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-Propenoate.

[0448] HCl (3 mL of a 4.0 M solution in 1,4-dioxane) was added to asolution ofethyl-3-[BOC-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate (0.220 g,0.273 mmol, 1 equiv) in 1,4-dioxane (3 mL). The reaction solution wasstirred at 23° C. for 2.25 hours and then concentrated to provide crudeethyl-3-[H₂N-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate•HCl whichwas set aside.

[0449] Cyclopentanol (0.431 mL, 4.75 mmol, 17.4 equiv) was dissolved indry CH₂Cl₂ (25 mL). Triethylamine (0.662 mL, 4.75 mmol, 17.4 equiv) andtriphosgene (0.507 g, 1.71 mmol, 6.26 equiv) were added sequentially.The reaction solution was stirred 2.5 hours to provide a stock solutionof cyclopentyl chloroformate (0.19 M). A portion of this solution (2.87mL, 0.545 mmol, 2 equiv) was added to a solution of crudeethyl-3-[H₂N-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate•HCl (fromabove) and 4-methylmorpholine (0.120 mL, 1.09 mmol, 4 equiv) in dryCH₂Cl₂ (4 mL). The reaction mixture was stirred for 1.75 hours, thenpoured into water (30 mL), and extracted with CH₂Cl₂ (3×30 mL). Thecombined organic phases were dried over Na₂SO₄, concentrated, and theresidue was chromatographed on silica gel (gradient elution 40→50% EtOAcin hexanes) to affordethyl-3-[CyPentylOCO-L-ValΨ[COCH2]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.077 g, 35%) as a colorless glass: ¹H NGR (CDCl₃) δ 0.68 (d, 3H,J=6.8), 0.93 (d, 3H, J=6.8), 1.29 (t, 3H, J=7.2), 1.50-1.86 (m, 10H),1.93-2.04 (m, 2H), 2.29-2.38 (m, 2H), 2.52 (d, 1H, J=15.9), 2.64 (dd,1H, J=12.1, 5.6), 2.73-2.90 (m, 2H), 2.99 (dd, 1H, J=17.0, 9.8),4.10-4.22 (m, 3H), 4.40-4.51 (m, 1H), 4.92 (d, 1H, J=8.1), 4.95-5.03 (m,1H), 5.38 (d, 1H, J=15.9), 6.06 (d, 1H, J=8.4), 6.60 (dd, 1H, J=15.9,4.8), 6.92-7.00 (m, 2H), 7.04-7.11 (m, 2H), 7.16-7.32 (m, 15H).

[0450] Preparation of ProductEthyl-3-(CyPentylOCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-Gln)-E-Propenoate.

[0451]Ethyl-3-119-[CyPentylOCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-(Tr-Gln)]-E-propenoate(0.076 g, 0.093 mmol, 1 equiv) was dissolved in dry CH₂Cl₂ (6 mL).Triisopropylsilane (0.057 mL, 0.278 mmol, 3 equiv) and trifluoroaceticacid (3 mL) were added sequentially producing a bright yellow solution.This solution was stirred for 40 minutes and then concentrated. Theresidue was stirred in Et₂O (6 mL), and the solid was collected byfiltration, washed with Et₂O (2×3 mL), and then dried under vacuum togive ethyl-3-(CyPentylOCO-L-ValΨ[COCH₂]-L-(p-F)Phe-L-Gln)-E-propenoate(0.040 g, 75%) as a white solid: mp=220-222° C. (dec); R_(f)=0.16 (5%CH₃OH in CH₂Cl₂); IR (cm⁻¹) 3413, 3317, 1708, 1658; ¹H NGR (DMSO-d₆) δ0.74 (d, 3H, J=6.5), 0.82 (d, 3H, J=6.5), 1.20 (t, 3H, J=7.2), 1.38-1.84(m, 10H), 1.96-2.12 (m, 3H), 2.46-2.84 (m, 4H), 2.88-2.97 (m, 1H),3.82-3.89 (m, 1H), 4.09 (q, 2H, J=7.2), 4.25-4.36 (m, 1H), 4.84-4.94 (m,1H), 5.40 (d, 1H, J=15.6), 6.61 (dd, 1H, J=15.6, 5.3), 6.73 (s, 1H),6.98-7.24 (m, 5H), 7.30 (d, 1H, J=8.7), 7.99 (d, 1H, J=8.4); Anal.(C₃₀H₄₂FN₃O₇.0.5 H₂O) C, H, N.

Biochemical and Biological Evaluation

[0452] Inhibition of Rhinovirus Protease

[0453] Stock solutions (50 mM, in DMSO) of various compounds wereprepared; dilutions were in the same solvent. Recombinant Rhinovirus 3Cproteases from serotypes 14, 16, and 2 were prepared by the followingstandard chromatographic procedures: (1) ion exchange using Q SepharoseFast Flow from Pharmacia; (2) affinity chromatography using Affi-GelBlue from Biorad; and (3) sizing using Sephadex G-100 from Pharmacia.Assays contained 2% DMSO, 50 mM tris pH 7.6, 1 mM EDTA, a compound atthe indicated concentrations, approximately 1 μM substrate, and 50-100nM protease. For K_(i) determinations, the compound and werepreincubated for 10 minutes at 30° C. prior to addition of the substratestart). The k_(obs/I) values were obtained from reactions initiated byaddition of enzyme rather than substrate. RVP activity is measured inthe fluorescence resonance energy transfer assay. The substrate was(N-terminal) DABCYL Gly-Arg-Ala-Val-Phe-Gln-Gly-Pro-Val-Gly)-EDANS. Inthe uncleavaed peptide, the EDANS fluorescence was quenched by theproximal DABCYL moiety. When the peptide was cleaved, the quenching wasrelieved, and activity was measured as an increase in fluorescencesignal. Data was analyzed using standard non-linear fitting programs(Enzfit), and are shown in Table 1. TABLE 1 COMPOUND RVP INHIB k_(obs/I)(M⁻¹sec⁻¹)  1 ND 17,380 (2) ND 2,242 (16) ND 3,880  2 ND 47,000 (2) ND4,600 (16) ND 10,410  3 >7 μM(K_(i)) 29,200  4 ND 180,000 (2) ND 17,800(16) ND 34,600  5 ND 500,000 (2) ND 26,900 (16) ND 89,700  6 ND 87,600(2) ND 13,350 (16) ND 23,230  7 ND 255,000 (2) ND 25,000 (16) ND 100,000 8 ND 55,700 (2) ND 7,000 (16) ND 14,200 1 + 9 (˜1:1) ND 5,100 10 ND440,000 11 ND 850,000 12 ND 404,000 13 1.6 196,000 14 1.7 293,000 15 ND127,000 16 3.4 150,000 17 ND 845,000 18 0.78 127,400 19 ND 67,200 20 ND52,140 21 ND 243,000 22 ND 124,000 23 ND 36,500 24 0.67 240,000 25 ND16,000

[0454] In the above table, all data are for RVP serotype-14 unlessotherwise noted in parentheses. All strains of human rhinovirus (HRV)were purchased from American Type Culture Collection (ATCC) except forserotype 14, which was produced from the infectious cDNA cloneconstructed and supplied to Applicants by Dr. Robert Rueckert at theInstitute for Molecular Virology, University of Wisconsin, Madison, Wis.The column designated INHIB represents the percent inhibition at 10minute preincubation with 50 nM RVP prior to addition of substrate. Thedata in the column designated k_(obs/I) was measured from progresscurves enzyme start experiments. The designation ND indicates that avalue was not determined for that compound.

[0455] Antirhinoviral HI-HeLa Cell Culture Assay

[0456] In the Cell Protection Assay, the ability of compounds to protectcells against HRV infection was measured by the XTT dye reductionmethod. This method is described in Weislow, O. S., Kiser, R., Fine, D.L., Bader, J., Shoemaker, R. H., Boyd, M. R., J. Natl. Cancer Inst.1989, 81, 577-586, the disclosure of which is incorporated herein byreference.

[0457] HI-HeLa cells were infected with HRV-14 at a multiplicity ofinfection (m.o.i.) of 0.13 (virus particles/cell) or mock-infected withmedium only. Infected or mock-infected cells were resuspended at 8×10⁵cells per mL and incubated with appropriate concentrations of compoundsof formula I. Two days later, XTT/PMS was added to test plates, and theamount of formazan produced was quantified spectrophotometrically at450/650 nm. The EC₅₀ was calculated as the concentration of compoundthat increased the percentage of formazan production incompound-treated, virus infected cells to 50% of that produced bycompound-free mock-infected cells. The 50% cytotoxic dose (CC₅₀) wascalculated as the concentration of compound that decreased thepercentage of formazan produced in compound-treated, mock-infected cellsto 50% of that produced by compound-free mock-infected cells. Thetherapeutic index (TI) was calculated by dividing the CC₅₀ by the EC₅₀.

[0458] All strains of human rhinovirus (HRV) for use in this assay werepurchased from American Type Culture Collection (ATCC) except for HRVserotype-14, which was produced from the infectious cDNA cloneconstructed and supplied to Applicants by Dr. Robert Rueckert at theInstitute for Molecular Virology, University of Wisconsin, Madison, Wis.HRV stocks were propagated, and viral assays were performed in HI-HeLacells (ATCC). Cells were grown in Minimal Essential Medium, availablefrom Life Technologies, with 10% fetal bovine serum.

[0459] The compounds were tested against control compounds WIN 51711,WIN 52084, and WIN 54954, all obtained from Sterling-WinthropPharmaceuticals, and control compound Pirodavir, obtained from JanssenPharmaceuticals. Antiviral data obtained for the test compounds areshown in Table 2 where all data are for HRV serotype-14 unless otherwisenoted in parentheses. TABLE 2 EC₅₀ CC₅₀ # HRV (μM) (μM) TI  10.36 >320 >889  2 0.24 >320 >1333 (2) 1.8 >320 >178  3 1.9 50.1 26  40.10 >320 >3200 (2) 0.50 >320 >640  5 0.19 >320 >1730  6 0.68 >100 >147 7 0.022 >10 >454 (2) 0.10 >10 >100 (10) 0.035 >10 >286 (89)0.004 >10 >2500 (39) 0.13 >10 >75  8 0.19 >100 >526 1 + 9 1.3 >320 >246(˜1:1) (16) 2.8 >320 >114 (2) 2.0 >320 >160 (10) 4.1 >320 >78 (89)5.1 >320 >63 10 0.011 >1 >91 (2) 0.18 >1 >57 11 0.006 >3 >500 (2)0.12 >3 >25 (39) 0.13 >3 >23 (10) 0.060 >3 >50 (16) 0.025 >3 >120 (1A)0.16 >3 >18 12 0.14 >1 >7 13 0.060 >3 >50 14 0.020 >10 >500 (2)0.13 >10 >76 (89) 0.080 >10 >125 (16) 0.10 >10 >100 (10) 0.16 >10 >62(1A) 0.17 >10 >58 (39) 0.07 >10 >143 15 0.063 >1 >15 16 0.050 >3 >60 (2)0.18 >3 >16 (39) 0.20 >3 >15 (89) 0.080 >3 >37 (16) 0.10 >3 >30 (10)0.15 >3 >20 (1A) 0.18 >3 >16 17 0.027 >10 >370 (2) 0.36 >10 >27 (39)0.48 >10 >20 18 0.48 >3 >6 19 0.28 >3 >10 (2) 0.71 >3 >4 (10)1.6 >3 >1.8 (1A) 0.60 >3 >5 20 0.042 >3 >71 (2) 0.56 >3 >5 (39)1.2 >3 >2.5 (89) 0.47 >3 >6 (16) 0.15 >3 >20 (10) 0.50 >3 >6 (1A)0.53 >3 >5 21 0.16 >10 >62 (2) 1.0 >10 >10 (10) 1.4 >10 >7 (1A)0.56 >10 >17 22 0.050 >32 >640 (2) 0.36 >32 >88 (39) 0.40 >32 >80 (89)0.24 >32 >133 (16) 0.40 >32 >80 (10) 0.50 >32 >64 (1A) 0.43 >32 >74 2320.9 >10 ND 24 0.032 >3 >93 25 0.14 >3 >21 (2) 0.68 >3 >4 (10)1.7 >3 >1.7 (1A) 0.90 >3 >3 (16) 0.71 >3 >4 (39) 0.56 >3 >5 (89)0.32 >3 >9 WIN 0.78 >60 >77 51711 WIN 0.07 >10 >143 52084 WIN2.13 >63 >30 54954 Pirodavir 0.03 >10 >300

[0460] Anticoxsackieviral HI-HeLa Cell Culture Assay

[0461] The Coxsackie strain A-21 (CVA-21) was purchased from AmericanType Culture Collection (ATCC). Virus stocks were propagated, andantiviral assays were performed in HI-HeLa cells (ATCC). Cells weregrown in Minimal Essential Medium with 10% fetal bovine serum.

[0462] The ability of compounds to protect cells against CVA-21infection was measured by the XTT dye reduction method. This method isdescribed in Weislow, O. S., Kiser, R., Fine, D. L., Bader, J.,Shoemaker, R. H., Boyd, M. R., J. Natl. Cancer Inst. 1989, 81, 577-586,the disclosure of which is incorporated herein by reference. HI-HeLacells were infected with CVA-21 at a multiplicity of infection (m.o.i.)of 0.05 (CVA-21) or mock-infected with medium only. Infected oruninfected cells were resuspended at 4×10⁴ cells per mL and incubatedwith appropriate concentrations of drug. One day later, XTT/PMS wasadded to test plates, and the amount of formazan produced was quantifiedspectrophotometrically at 450/650 nm. The EC₅₀ was calculated as theconcentration of drug that increased the percentage of formazanproduction in drug-treated, virus-infected cells to 50% of that producedby drug-free, uninfected cells. The 50% cytotoxic dose (CC₅₀) wascalculated as the concentration of drug that decreased the percentage offormazan produced in drug-treated, uninfected cells to 50% of thatproduced in drug-free, uninfected cells. The therapeutic index (TI) wascalculated by dividing the CC₅₀ by the EC₅₀.

[0463] The compounds were tested against control compound WIN 54954,obtained from Sterling-Winthrop Pharmaceuticals, and control compoundPirodavir, obtained from Janssen Pharmaceuticals. Antiviral dataobtained for the test compounds against Coxsackie strain A-21 (CVA-21)are shown in Table 3. TABLE 3 COMPOUND # EC₅₀ (μM) CC₅₀ (μM) TI 70.16 >10 >63 WIN 54954 >100 >100 Pirodavir >100 >100

[0464] In describing the invention, the inventors have set forth certaintheories and mechanisms in an effort to disclose how or why theinvention works in the manner in which it works. These theories andmechanisms are set forth for informational purposes only. Applicants arenot to be bound by any specific chemical or physical mechanisms ortheories of operation.

[0465] While the invention has been described in terms of variouspreferred embodiments and specific examples, those skilled in the artwill recognize that various changes and modifications can be madewithout departing from the spirit and scope of the invention, as definedin the appended claims. TABLE 1 COMPOUND RVP INHIB k_(obs/I) (M⁻¹ sec⁻¹) 1 ND 17,380 (2) ND 2,242 (16) ND 3,880  2 ND 47,000 (2) ND 4,600 (16)ND 10,410  3 >7 μM(K_(i)) 29,200  4 ND 180,000 (2) ND 17,800 (16) ND34,600  5 ND 500,000 (2) ND 26,900 (16) ND 89,700  6 ND 87,600 (2) ND13,350 (16) ND 23,230  7 ND 255,000 (2) ND 25,000 (16) ND 100,000  8 ND55,700 (2) ND 7,000 (16) ND 14,200 1 + 9 (˜1:1) ND 5,100 10 ND 440,00011 ND 850,000 12 ND 404,000 13 1.6 196,000 14 1.7 293,000 15 ND 127,00016 3.4 150,000 17 ND 845,000 18 0.78 127,400 19 ND 67,200 20 ND 52,14021 ND 243,000 22 ND 124,000 23 ND 36,500 24 0.67 240,000 25 ND 16,000

[0466] In the above table, all data are for RVP serotype-14 unlessotherwise noted in parentheses. All strains of human rhinovirus (HRV)were purchased from American Type Culture Collection (ATCC) except forserotype 14, which was produced from the infectious cDNA cloneconstructed and supplied to Applicants by Dr. Robert Rueckert at theInstitute for Molecular Virology, University of Wisconsin, Madison, Wis.The column designated INHIB represents the percent inhibition at 10minute preincubation with 50 nM RVP prior to addition of substrate. Thedata in the column designated k_(obs/I) was measured from progresscurves in enzyme start experiments. The designation ND indicates that avalue was not determined for that compound.

[0467] Antirhinoviral HI-HeLa Cell Culture Assay

[0468] In the Cell Protection Assay, the ability of compounds to protectcells against HRV infection was measured by the XTT dye reductionmethod. This method described in Weislow, O. S., Kiser, R., Fine, D. L.,Bader, J., Shoemaker, R. H., Boyd, M. R., J. Natl. Cancer Inst. 1989,81, 577-586, the disclosure of which is incorporated herein byreference.

[0469] HI-HeLa cells were infected with HRV-14 at a multiplicity ofinfection (m.o.i.) of 0.13 (virus particles/cell) or mock-infected withmedium only. Infected or mock-infected cells were resuspended at 8×10⁵cells per mL and incubated with appropriate concentrations of compoundsof formula I. Two days later, XTT/PMS was added to test plates, and theamount of formazan produced was quantified spectrophotometrically at450/650 nm. The EC₅₀ was calculated as the concentration of compoundthat increased the percentage of formazan production incompound-treated, virus infected cells to 50% of that produced bycompound-free mock-infected cells. The 50% cytotoxic dose (CC₅₀) wascalculated as the concentration of compound that decreased thepercentage of formazan produced in compound-treated, mock-infected cellsto 50% of that produced by compound-free mock-infected cells. Thetherapeutic index (TI) was calculated by dividing the CC₅₀ by the EC₅₀.

[0470] All strains of human rhinovirus (HRV) for use in this assay werepurchased from American Type Culture Collection (ATCC) except for HRVserotype-14, which was produced from the infectious cDNA cloneconstructed and supplied to Applicants by Dr. Robert Rueckert at theInstitute for Molecular Virology, University of Wisconsin, Madison, Wis.HRV stocks were propagated, and viral assays were performed in HI-HeLacells (ATCC). Cells were grown in Minimal Essential Medium, availablefrom Life Technologies, with 10% fetal bovine serum.

[0471] The compounds were tested against control compounds WIN 51711,WIN 52084, and WIN 54954, all obtained from Sterling-WinthropPharmaceuticals, and control compound Pirodavir, obtained from JanssenPharmaceuticals. Antiviral data obtained for the test compounds areshown in Table 2 where all data are for HRV serotype-14 unless otherwisenoted in parentheses. TABLE 2 EC₅₀ CC₅₀ # HRV (μM) (μM) TI  10.36 >320 >889  2 0.24 >320 >1333 (2) 1.8 >320 >178  3 1.9 50.1 26  40.10 >320 >3200 (2) 0.50 >320 >640  5 0.19 >320 >1730  6 0.68 >100 >147 7 0.022 >10 >454 (2) 0.10 >10 >100 (10) 0.035 >10 >286 (89)0.004 >10 >2500 (39) 0.13 >10 >75  8 0.19 >100 >526 1 + 9 1.3 >320 >246(˜1:1) (16) 2.8 >320 >114 (2) 2.0 >320 >160 (10) 4.1 >320 >78 (89)5.1 >320 >63 10 0.011 >1 >91 (2) 0.18 >1 >57 11 0.006 >3 >500 (2)0.12 >3 >25 (39) 0.13 >3 >23 (10) 0.060 >3 >50 (16) 0.025 >3 >120 (1A)0.16 >3 >18 12 0.14 >1 >7 13 0.060 >3 >50 14 0.020 >10 >500 (2)0.13 >10 >76 (89) 0.080 >10 >125 (16) 0.10 >10 >100 (10) 0.16 >10 >62(1A) 0.17 >10 >58 (39) 0.07 >10 >143 15 0.063 >1 >15 16 0.050 >3 >60 (2)0.18 >3 >16 (39) 0.20 >3 >15 (89) 0.080 >3 >37 (16) 0.10 >3 >30 (10)0.15 >3 >20 (1A) 0.18 >3 >16 17 0.027 >10 >370 (2) 0.36 >10 >27 (39)0.48 >10 >20 18 0.48 >3 >6 19 0.28 >3 >10 (2) 0.71 >3 >4 (10)1.6 >3 >1.8 (1A) 0.60 >3 >5 20 0.042 >3 >71 (2) 0.56 >3 >5 (39)1.2 >3 >2.5 (89) 0.47 >3 >6 (16) 0.15 >3 >20 (10) 0.50 >3 >6 (1A)0.53 >3 >5 21 0.16 >10 >62 (2) 1.0 >10 >10 (10) 1.4 >10 >7 (1A)0.56 >10 >17 22 0.050 >32 >640 (2) 0.36 >32 >88 (39) 0.40 >32 >80 (89)0.24 >32 >133 (16) 0.40 >32 >80 (10) 0.50 >32 >64 (1A) 0.43 >32 >74 2320.9 >10 ND 24 0.032 >3 >93 25 0.14 >3 >21 (2) 0.68 >3 >4 (10)1.7 >3 >1.7 (1A) 0.90 >3 >3 (16) 0.71 >3 >4 (39) 0.56 >3 >5 (89)0.32 >3 >9 WIN 0.78 >60 >77 51711 WIN 0.07 >10 >143 52084 WIN2.13 >63 >30 54954 Pirodavir 0.03 >10 >300

[0472] Anticoxsackieviral HI-HeLa Cell Culture Assay

[0473] The Coxsackie strain A-21 (CVA-21) was purchased from AmericanType Culture Collection (ATCC). Virus stocks were propagated, andantiviral assays were performed in HI-HeLa cells (ATCC). Cells weregrown in Minimal Essential Medium with 10% fetal bovine serum.

[0474] The ability of compounds to protect cells against CVA-21infection was measured by the XTT dye reduction method. This method isdescribed in Weislow, O. S., Kiser, R., Fine, D. L., Bader, J.,Shoemaker, R. H., Boyd, M. R., J. Natl. Cancer Inst. 1989, 81, 577-586,the disclosure of which is incorporated herein by reference. HI-HeLacells were infected with CVA-21 at a multiplicity of infection (m.o.i.)of 0.05 (CVA-21) or mock-infected with medium only. Infected oruninfected cells were resuspended at 4×10⁴ cells per mL and incubatedwith appropriate concentrations of drug. One day later, XTT/PMS wasadded to test plates, and the amount of formazan produced was quantifiedspectrophotometrically at 450/650 nm. The EC₅₀ was calculated as theconcentration of drug that increased the percentage of formazanproduction in drug-treated, virus-infected cells to 50% of that producedby drug-free, uninfected cells. The 50% cytotoxic dose (CC₅₀) wascalculated as the concentration of drug that decreased the percentage offormazan produced in drug-treated, uninfected cells to 50% of thatproduced in drug-free, uninfected cells. The therapeutic index (TI) wascalculated by dividing the CC₅₀ by the EC₅₀.

[0475] The compounds were tested against control compound WIN 54954,obtained from Sterling-Winthrop Pharmaceuticals, and control compoundPirodavir, obtained from Janssen Pharmaceuticals. Antiviral dataobtained for the test compounds against Coxsackie strain A-21 (CVA-21)are shown in Table 3. TABLE 3 COMPOUND # EC₅₀ (μM) CC₅₀ (μM) TI 70.16 >10 >63 WIN 54954 >100 >100 Pirodavir >100 >100

[0476] In describing the invention, the inventors have set forth certaintheories and mechanisms in an effort to disclose how or why theinvention works in the manner in which it works. These theories andmechanisms are set forth for informational purposes only. Applicants arenot to be bound by any specific chemical or physical mechanisms ortheories of operation.

[0477] While the invention has been described in terms of variouspreferred embodiments and specific examples, those skilled in the artwill recognize that various changes and modifications can be madewithout departing from the spirit and scope of the invention, as definedin the appended claims.

What is claim is:
 1. At least one compound of the formula (I):

wherein M is O or S; R₁ is H, F, an alkyl group, OH, SH, or an O-alkylgroup; R₂ and R₅ are independently selected from H,

or an alkyl group, wherein said alkyl group is different from

with the proviso that at least one of R₂ or R₅ must be

and wherein, when R₂ or R₅ is

X is ═CH or ═CF and Y₁ is ═CH or ═CF, or X and Y₁ together with Q′ forma three-membered ring in which Q′ is —C(R₁₀)(R₁₁)— or —O—, X is —CH— or—CF—, and Y₁ is —CH—, —CF—, or —C(alkyl)-, where R₁₀ and R₁₁independently are H, a halogen, or an alkyl group, or, together with thecarbon atom to which they are attached, form a cycloalkyl group or aheterocycloalkyl group, or X is —CH₂—, —CF₂—, —CHF—, or —S—, and Y₁ is—O—, —S—, —NR₁₂—, —C(R₁₃)(R₁₄)—, —C(O)—, —C(S)—, or —C(CR₁₃R₁₄)—,wherein R₁₂ is H or alkyl, and R₁₃ and R₁₄ independently are H, F, or analkyl group, or, together with the atoms to which they are bonded, forma cycloalkyl group or a heterocycloalkyl group; A₁is C, CH, CF, S, P,Se, N, NR₁₅, S(O), Se(O), P—OR₁₅, or P—NR₁₅R₁₆, wherein R₁₅ and R₁₆independently are an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, or a heteroaryl group, or, together with the atomto which they are bonded, form a heterocycloalkyl group; D₁ is a moietywith a lone pair of electrons capable of forming a hydrogen bond; and B₁is H, F, an alkyl group, a cycloalkyl group, a heterocycloalkyl group,an aryl group, a heteroaryl group, —OR₁₇, —SR₁₇, —NR₁₇R₁₈, —NR₁₉NR₁₇R₁₈,or —NR₁₇OR₁₈, wherein R₁₇, R₁₈, and R₁₉ independently are H, an alkylgroup, a cycloalkyl group, a heterocycloalkyl group, an aryl group, aheteroaryl group, or an acyl group; and with the provisos that when D₁is the moiety ≡N with a lone pair of electrons capable of forming ahydrogen bond, B₁ does not exist; and when A₁ is an sp³ carbon, B₁ isnot —NR₁₇R₁₈ when D₁ is the moiety —NR₂₅R₂₆ with a lone pair ofelectrons capable of forming a hydrogen bond, wherein R₂₅ and R₂₆ areindependently H, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, or a heteroaryl group; and wherein D₁—A₁—B₁optionally forms a nitro group where A₁ is N; and further wherein, whenR₂ or R₅ is

X is ═CH or ═CF and Y₂ is ═C, ═CH, or ═CF, or X and Y₂ together with Q′form a three-membered ring in which Q′ is —C(R₁₀)(R₁₁)— or —O—, X is—CH— or —CF—, and Y₂ is —CH—, —CF—, or —C(alkyl)-, where R₁₀ and R₁₁,independently are H, a halogen, or an alkyl group, or, together with thecarbon atom to which they are attached, form a cycloalkyl group or aheterocycloalkyl group, or X is —CH₂—, —CF₂—, —CHF—, or —S—, and Y₂ is—O—, —S—, —N(R′₁₂)—, —C(O)—, —C(R′₁₃)(R′₁₄)—, —C(S)—, or —C(CR′₁₃R′₁₄)—,wherein R′₁₂ is H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —OR′₁₃,—NR′₁₃R′₁₄, —C(O)—R′₁₃, —SO₂R′₁₃, or —C(S)R′₁₃, and R′₁₃ and R′₁₄,independently are H, F, or an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, or a heteroaryl group, or,together with the atom to which they are attached, form a cycloalkylgroup or a heterocycloalkyl group; A₂ is C, CH, CF, S, P, Se, N, NR₁₅,S(O), Se(O), P—OR₁₅, or P—NR₁₅R₁₆, wherein R₁₅ and R₁₆ independently arean alkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, or a heteroaryl group, or, together with the atom to which theyare bonded, form a heterocycloalkyl group; D₂ is a moiety with a lonepair of electrons capable of forming a hydrogen bond; and B₂ is H, F, analkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, a heteroaryl group, —OR₁₇, —SR₁₇, —NR₁₇R₁₈, —NR₁₉NR₁₇R₁₈, or—NR₁₇OR₁₈, wherein R₁₇, R₁₈, and R₁₉ independently are H, an alkylgroup, a cycloalkyl group, a heterocycloalkyl group, an aryl group, aheteroaryl group, or an acyl group; and further wherein any combinationof Y₂, A₂, B₂, and D₂ optionally can form a cycloalkyl group, aheterocycloalkyl group, an aryl group, or a heteroaryl group; R₃ and R₆are independently H, F, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —C(O)R₁₇,—OR₁₇, —SR₁₇, —NR₁₇R₁₈, —NR₁₉NR₁₇R₁₈, or —NR₁₇OR₁₈, wherein R₁₇, R₁₈,and R₁₉ independently are H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, or an acylgroup; or, R₃ and R₆, together with the carbon atom to which they areattached, form a cycloalkyl group or a heterocycloalkyl group; R₇ is H,an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, a heteroaryl group, —OR₁₇, —SR₁₇, —NR₁₇R₁₈, —NR₁₉NR₁₇R₁₈, or—NR₁₇OR₁₈, wherein R₁₇, R₁₈, and R₁₉ independently are H, an alkylgroup, a cycloalkyl group, a heterocycloalkyl group, an aryl group, aheteroaryl group, or an acyl group; or R₇, together with R₃ or R₆ andthe atoms to which they are attached, forms a heterocycloalkyl group;R₂₀ is H, OH, or any suitable organic moiety; and Z and Z₁ areindependently H, F, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —C(O)R₂₁,—CO₂R₂₁, —CN, —C(O)NR₂₁,R₂₂, —C(O)NR₂₁OR₂₂, —C(S)R₂₁, —C(S)NR₂₁R₂₂,—NO₂, —SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂, —SO(NR₂₁)(OR₂₂), —SONR₂₁, —SO₃R₂₁,—PO(OR₂₁)₂, —PO(R₂₁)(R₂₂), —PO(NR₂₁R₂₂)(OR₂₃), PO(NR₂₁R₂₂)(NR₂₃R₂₄),—C(O)NR₂₁NR₂₂R₂₃, or —C(S)NR₂₁NR₂₂R₂₃, wherein R₂₁, R₂₂, R₂₃, and R₂₄are independently H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, an acylgroup, or a thioacyl group, or wherein any two of R₂₁, R₂₂, R₂₃, andR₂₄, together with the atom(s) to which they are bonded, form aheterocycloalkyl group; or Z₁, as defined above, together with R₁, asdefined above, and the atoms to which Z₁ and R₁ are bonded, form acycloalkyl or heterocycloalkyl group, or Z and Z₁, both as definedabove, together with the atoms to which they are bonded, form acycloalkyl or heterocycloalkyl group; or a pharmaceutically acceptableprodrug, salt, active metabolite, or solvate thereof; and wherein saidcompound, or pharmaceutically acceptable prodrug, salt, activemetabolite, or solvate thereof, has antipicomaviral activity with anEC₅₀ less than or equal to 10 μM in the HI-HeLa cell culture assay. 2.At least one compound of claim 1, wherein R₁ is H or F, or apharmaceutically acceptable prodrug, salt, active metabolite, or solvatethereof.
 3. At least one compound of claim 1, wherein R₂₀ is H, an alkylgroup, a cycloalkyl group, a heterocycloalkyl group, an aryl group, aheteroaryl group, —OR₁₇, —SR₁₇, —NR₁₇R₁₈, —NR₁₉NR₁₇R₁₈, or —NR₁₇OR₁₈,wherein R₁₇, R₁₈, and R₁₉ independently are H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, or an acyl group, or a pharmaceutically acceptable prodrug, salt,active metabolite, or solvate thereof.
 4. At least one compound of claim3, wherein R₂₀ is the alkyl group —C(R₄₁)(R₄₂)NR₄₃R₄₄, wherein: R₄₁, andR₄₂ independently are H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, or a heteroaryl group; and R₄₃and R₄₄ independently are H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —NR₄₅R₄₆,—C(O)R₄₅, —C(S)R₄₅, —C(O)NR₄₅R₄₆, —C(S)NR₄₅R₄₆, —C(O)NR₄₅OR₄₆,—C(S)NR₄₅OR₄₆, —C(O)SR₄₅, —C(O)OR₄₅, —C(S)OR₄₅, —C(S)SR₄₅, —OR₄₅, —SR₄₅,—C(O)NR₄₅NR₄₆R₄₇, —C(S)NR₄₅NR₄₆R₄₇, —SOR₄₅, —SO₂R₄₅, —S(O)NR₄₅R₄₆,—S(O)NR₄₅(OR₄₆), —SO₂NR₄₅R₄₆, —SO₂NR₄₅(OR₄₆), or —SO₃R₄₅, wherein R₄₅,R₄₆, and R₄₇ independently are H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, or whereinany suitable combination of R₄₁, R₄₂, R₄₃, and R₄₄ together form acycloalkyl group or a heterocycloalkyl group; or a pharmaceuticallyacceptable prodrug, salt, active metabolite, or solvate thereof.
 5. Atleast one compound of claim 4, wherein at least one of R₄₃ or R₄₄ is—C(O)SR₄₅ or —C(O)OR₄₅, or a pharmaceutically acceptable prodrug, salt,active metabolite, or solvate thereof.
 6. At least one compound of claim5, wherein R₄₅ is an alkyl group, a cycloalkyl group, an aryl group, aheterocycloalkyl group, or a heteroaryl group, or a pharmaceuticallyacceptable prodrug, salt, active metabolite, or solvate thereof.
 7. Atleast one compound of claim 6, wherein R₄₅ is a C₁₋C₁₀ alkyl group or acycloalkyl group, or a pharmaceutically acceptable prodrug, salt, activemetabolite, or solvate thereof.
 8. At least one compound of claim 1,wherein at least one of R₂ or R₅ is

or a pharmaceutically acceptable prodrug, salt, active metabolite, orsolvate thereof.
 9. At least one compound according to claim 8, whereinD₁ is —OR₂₅, ═O, ═S, ≡N, ═NR₂₅, or —NR₂₅R₂₆, wherein R₂₅ and R₂₆ areindependently H, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, or a heteroaryl group, or, together with thenitrogen atom to which they are bonded, form a heterocycloalkyl group;or a pharmaceutically acceptable prodrug, salt, active metabolite, orsolvate thereof.
 10. At least one compound according to claim 9, whereinD₁ is ═O; or a pharmaceutically acceptable prodrug, salt, activemetabolite, or solvate thereof.
 11. At least one compound according toclaim 8, wherein A₁ is C, CH, S, or S(O); or a pharmaceuticallyacceptable prodrug, salt, active metabolite, or solvate thereof.
 12. Atleast one compound according to claim 11, wherein A₁ is C; or apharmaceutically acceptable prodrug, salt, active metabolite, or solvatethereof.
 13. At least one compound according to claim 8, wherein B₁ isNR₁₇R₁₈, wherein R₁₇ and R₁₈ are independently H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, an acyl group; or a pharmaceutically acceptable prodrug, salt,active metabolite, or solvate thereof.
 14. At least one compoundaccording to claim 1, wherein at least one of R₂ or R₅

or a pharmaceutically acceptable prodrug, salt, active metabolite, orsolvate thereof.
 15. At least one compound according to claim 14,wherein D₂ is —OR₂₅, ═O, ═S, ≡N, ═NR₂₅, or —NR₂₅R₂₆, wherein R₂₅ and R₂₆are independently H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, or a heteroaryl group, or,together with the atom(s) to which they are bonded, form aheterocycloalkyl group; or a pharmaceutically acceptable prodrug, salt,active metabolite, or solvate thereof.
 16. At least one compoundaccording to claim 15, wherein D₂ is ═O; or a pharmaceuticallyacceptable prodrug, salt, active metabolite, or solvate thereof.
 17. Atleast one compound according to claim 14, wherein A₂ is C, CH, S, orS(O); or a pharmaceutically acceptable prodrug, salt, active metabolite,or solvate thereof.
 18. At least one compound according to claim 17,wherein A₂ is C; or a pharmaceutically acceptable prodrug, salt, activemetabolite, or solvate thereof.
 19. At least one compound according toclaim 14, wherein B₂ is —NR₁₇R₁₈, wherein R₁₇ and R₁₈ are independentlyH, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, a heteroaryl group, an acyl group; or a pharmaceuticallyacceptable prodrug, salt, active metabolite, or solvate thereof.
 20. Atleast one compound according to claim 1, wherein A₁ is C, CH, S, or S(O)or wherein A₂ is C, CH, S, or S(O); or a pharmaceutically acceptableprodrug, salt, active metabolite, or solvate thereof.
 21. At least onecompound according to claim 1, wherein Z and Z₁ are independently H, anaryl group, or a heteroaryl group, —C(O)R₂₁, —CO₂R₂₁, —CN,—C(O)NR₂₁,R₂₂, —C(O)NR₂₁OR₂₂, —C(S)R₂₁, —C(S)NR₂₁R₂₂, —NO₂, —SOR₂₁,—SO₂R₂₁, —SO₂NR₂₁R₂₂, —SO(NR₂₁)(OR₂₂), —SONR₂₁, —SO₃R₂₁,—C(O)NR₂₁NR₂₂R₂₃, or —C(S)NR₂₁NR₂₂R₂₃; wherein R₂₁, R₂₂, and R₂₃ areindependently H, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, a heteroaryl group, an acyl group, or a thioacylgroup, or wherein any two of R₂, R₂₂, and R₂₃, together with the atom(s)to which they are bonded, form a heterocycloalkyl group; or apharmaceutically acceptable prodrug, salt, active metabolite, or solvatethereof.
 22. At least one compound according to claim 1, wherein M is O.23. At least one compound having the formula X:

wherein R₆₁, is H, F, or an alkyl group; R₆₂ is selected from one of thefollowing moieties:

wherein R₃₅ is H, an alkyl group, an aryl group, —OR₃₈, or —NR₃₈R₃₉,wherein R₃₈ and R₃₉ independently are H, an alkyl group, a cycloalkylgroup, a heterocycloalkyl group, an aryl group, a heteroaryl group, oran acyl group; and R₃₆ is H or an alkyl group, or R₃₅ and R₃₆, togetherwith the nitrogen atom to which they are attached, form aheterocycloalkyl group or a heteroaryl group; R₃₇ is an alkyl group, anaryl group, or —NR₃₈R₃₉, wherein R₃₈ and R₃₉ are as defined above; R₅₀is H, an alkyl group, an aryl group, —OR₃₈, —SR₃₉, —NR₃₈R₃₉,—NR₄₀NR₃₈R₃₉, or —NR₃₈OR₃₉, or R₅₀and R₃₆, together with the atoms towhich they are attached, form a heterocycloalkyl group; wherein R₃₈ andR₃₉ are as defined above, and R₄₀ is H, an alkyl group, a cycloalkylgroup, a heterocycloalkyl group, an aryl group, a heteroaryl group, oran acyl group; and n is 0, 1, or2; R₆₃ is H or an alkyl group; R₆₄ is H,an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, or a heteroaryl group; R₆₅ is H or an alkyl group; R₆₆ is H, anacyl group, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, a sulfonyl group, or a heteroaryl group; R₆₇ is Hor an alkyl group; and Z and Z₁ are independently H, F, an alkyl group,a cycloalkyl group, a heterocycloalkyl group, an aryl group, aheteroaryl group, —C(O)R₂₁, —CO₂R₂₁, —CN, —C(O)NR₂₁R₂₂, —C(O)NR₂₁OR₂₂,—C(S)R₂₁, —C(S)NR₂₁R₂₂, —NO₂, —SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂,—SO(NR₂₁)(OR₂₂), —SONR₂₁, —SO₃R₂₁, —PO(OR₂₁)₂, —PO(R₂₁)(R₂₂),—PO(NR₂₁R₂₂)(OR₂₃), —PO(NR₂₁R₂₂)(NR₂₃R₂₄), —C(O)NR₂₁NR₂₂R₂₃, or—C(S)NR₂₁NR₂₂R₂₃, wherein R₂₁, R₂₂, R₂₃, and R₂₄ are independently H, analkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, a heteroaryl group, an acyl group, or a thioacyl group, orwherein any two of R₂₁, R₂₂, R₂₃, and R₂₄, together with the atom(s) towhich they are bonded, form a heterocycloalkyl group, or Z and Z₁, bothas defined above, together with the atoms to which they are bonded, forma heterocycloalkyl group; or a pharmaceutically acceptable prodrug,salt, active metabolite, or solvate thereof.
 24. At least one compoundaccording to claim 23, wherein R₆₆ is the acyl group —C(O)OR₆₈ or theacyl group —C(O)SR₆₈, wherein R₆₈ is an alkyl group, a cycloalkyl group,an aryl group, a heterocycloalkyl group, or a heteroaryl group, or apharmaceutically acceptable prodrug, salt, active metabolite, or solvatethereof.
 25. At least one compound according to claim 4, having theformula II:

wherein R₁, R₅, R₆, R₇, R₄₂, R₄₃, and Z are H, R₂ is CH₂CH₂C(O)NH₂, andR₃ is CH₂Ph, R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

R₃ is CH₂Ph, R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

R₃ is CH₂Ph, R₄₁ is CH₂CH(CH₃)₂, Z₁ is

 and R₄₄ is

R₃ is CH₂Ph, R₄₁ is CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

R₃ is CH₂Ph, R₄₁ is CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

R₃ is CH₂Ph, R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

R₃ is CH₂Ph, R₄₁ is CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

R₃ is CH₂Ph, R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

R₃ is CH₂(p-CH₃)Ph, R₄₁ is CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is R₃ isCH₂(p-CH₃)Ph, R₄₁ is CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

or a pharmaceutically acceptable prodrug, salt, active metabolite, orsolvate thereof.
 26. At least one compound according to claim 4, havingthe formula III:

wherein R₁, R₅, R₆, R₇, R₄₂, R₄₃, and Z are H, R₃ is CH₂Ph, R₂ isCH₂CH₂C(O)NH₂, R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

or a pharmaceutically acceptable prodrug, salt, active metabolite, orsolvate thereof.
 27. At least one compound of the formula (IV):

wherein: R₁, R₅, R₆, R₇, and R₄₂ are H, R₂ is CH₂CH₂C(O)NH₂, and R₃ isCH₂(p-CH₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄ is CH₂Ph, and R₄₄ is

R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH(CH₃)₂, and R₄₄ is

R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH(CH₃)₂, and R₄₄ is

R₃ is CH₂(p-CF₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH(CH₃)₂, and R₄₄

R₃ is CH₂(p-CF₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH(CH₃)₂, and R₄₄

R₃ is CH₂(p-CH₃)Ph, Z and Z₁ together form

 (where * indicates the point of attachment and the carbonyl group iscis to the R₁ group), R₄₁ is CH(CH₃)₂, and R₄₄ is

R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH₂Ph, and R₄₄ is

R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH₂CH(CH₃)₂, and R₄₄is

R₃ is CH₂(p-CH₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH(CH₃)₂, and R₄₄ is

R₃ is CH₂(p-CH₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH₂CH(CH₃)₂, and R₄₄is

R₃ is CH₂Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is C(CH₃)₃, and R₄₄ is

R₃ is CH₂(p-CH₃)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH(CH₃)₂, and R₄₄ is

R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is cyclohexyl, and R₄₄ is

 or R₃ is CH₂(p-F)Ph, Z is H, Z₁ is CO₂CH₂CH₃, R₄₁ is CH(CH₃)₂, and R₄₄is

or a pharmaceutically acceptable prodrug, salt, active metabolite, orsolvate thereof.
 28. A composition comprising at least one compound offormula II:

wherein R₁ , R₅, R₆, R₇, R₄₂, R₄₃, and Z are H, R₃ is CH₂Ph, R₂ isCH₂CH₂C(O)NH₂, R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

or a pharmaceutically acceptable prodrug, salt, active metabolite, orsolvate thereof, and at least one compound of formula III:

wherein R₁ , R₅, R₆, R₇, R₄₂, R₄₃, and Z are H, R₃ is CH₂Ph, R₂ isCH₂CH₂C(O)NH₂, R₄₁ is CH₂CH(CH₃)₂, Z₁ is CO₂CH₂CH₃, and R₄₄ is

or a pharmaceutically acceptable prodrug, salt, active metabolite, orsolvate thereof.
 29. A pharmaceutical composition comprising: (a) atherapeutically effective amount of at least one compound as defined inclaim 1 or a pharmaceutically acceptable prodrug, salt, activemetabolite, or solvate thereof; and (b) a pharmaceutically acceptablecarrier, diluent, vehicle, or excipient.
 30. A method of treating amammalian disease condition mediated by picornaviral protease activitythat comprises administering to a mammal in need thereof atherapeutically effective amount of at least one compound as defined inclaim 1 or a pharmaceutically acceptable prodrug, salt, activemetabolite, or solvate thereof.
 31. A method of inhibiting the activityof a picornaviral 3C protease that comprises contacting the picornaviral3C protease with an effective amount of at least one compound as definedin claim 1 or a pharmaceutically acceptable prodrug, salt, activemetabolite, or solvate thereof.
 32. A method of inhibiting the activityof a rhinoviral protease that comprises contacting the rhinoviralprotease with an effective amount of at least one compound as defined inclaim 1 or a pharmaceutically acceptable prodrug, salt, activemetabolite, or solvate thereof.
 33. A compound according to claim 1, ora pharmaceutically acceptable prodrug, salt, active metabolite, orsolvate thereof, wherein said antipicornaviral activity isantirhinoviral activity.
 34. A compound according to claim 1, or apharmaceutically acceptable prodrug, salt, active metabolite, or solvatethereof, wherein said antipicornaviral activity is anticoxsackieviralactivity.